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collecting pollen from bees

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  • Diane L Larson
    Hi all, Sam Droege and I will be starting a pollination network study at Badlands National Park next summer. As part of this study, we want to collect pollen
    Message 1 of 10 , Nov 10, 2009
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      Hi all,

      Sam Droege and I will be starting a pollination network study at Badlands National Park next summer.  As part of this study, we want to collect pollen carried by individual bees as an indication of which flowers they have been visiting prior to collection (see Bosch et al. 2009 Ecol. Letters 12: 409-418).  We have two questions.  First, is there an alternative to individual kill vials, given that we want insect-specific pollen information?  Working in the backcountry, it will be difficult to handle the number of vials we'll likely need.  Second, in removing pollen from the insects' bodies, is the method described in Kearns and Inouye 1993, i.e., rubbing a small cube of fuchsine stained gelatin over the insect's body, still the best method?  If so, how soon must this be done after capture?

      If anyone has experience with mutualist networks in general, I'd love to talk with you.

      Thanks for any advice!

      Diane
      *******************************************
      Diane L. Larson
      Research Biologist
      USGS Northern Prairie Wildlife Research Center
      1561 Lindig St.
      St. Paul, MN  55108

      Voice 651-649-5041
      FAX 651-649-5040
      Email: dlarson@...

      “It’s a poor sort of memory that only works backwards,” the Queen remarked.
      - Lewis Carroll, Through the Looking-Glass
    • David Inouye
      Pollen is pretty indestructible so there s no rush on getting the pollen off, unless the bees would be shaken around enough to remove pollen from their bodies.
      Message 2 of 10 , Nov 10, 2009
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        Pollen is pretty indestructible so there's no rush on getting the pollen off, unless the bees would be shaken around enough to remove pollen from their bodies.  Washing is another alternative. 

        David

        At 11:24 AM 11/10/2009, you wrote:
         


        Hi all,

        Sam Droege and I will be starting a pollination network study at Badlands National Park next summer.  As part of this study, we want to collect pollen carried by individual bees as an indication of which flowers they have been visiting prior to collection (see Bosch et al. 2009 Ecol. Letters 12: 409-418).  We have two questions.  First, is there an alternative to individual kill vials, given that we want insect-specific pollen information?  Working in the backcountry, it will be difficult to handle the number of vials we'll likely need.  Second, in removing pollen from the insects' bodies, is the method described in Kearns and Inouye 1993, i.e., rubbing a small cube of fuchsine stained gelatin over the insect's body, still the best method?  If so, how soon must this be done after capture?

        If anyone has experience with mutualist networks in general, I'd love to talk with you.

        Thanks for any advice!

        Diane
        *******************************************
        Diane L. Larson
        Research Biologist
        USGS Northern Prairie Wildlife Research Center
        1561 Lindig St.
        St. Paul, MN  55108

        Voice 651-649-5041
        FAX 651-649-5040
        Email: dlarson@...
      • Maria Stanko
        Hi Diane, I have collected this kind of data for my dissertation and use methods put together though talking with several other researchers who use similar
        Message 3 of 10 , Nov 10, 2009
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          Hi Diane,
          I have collected this kind of data for my dissertation and use methods put
          together though talking with several other researchers who use similar
          techniques. I bring about 10 collecting vials into the field and line
          them with paper baggies (nothing fancy, I roll and fold up squares of
          paper around the end of my finger). When they're filled, I close and
          label the bags and re-line the tubes. You've still got to carry all those
          baggies but it should be easier than a lot of vials. Or you could
          transfer specimens to little pill boxes.
          I use the fuchsin jelly method, the key is that you don't need a lot of
          jelly! The hardest part is learning to or getting someone to id the
          pollen.
          Feel free to email me off-list for more details,
          Maria Stanko

          > Hi all,
          >
          > Sam Droege and I will be starting a pollination network study at Badlands
          > National Park next summer. As part of this study, we want to collect
          > pollen carried by individual bees as an indication of which flowers they
          > have been visiting prior to collection (see Bosch et al. 2009 Ecol.
          > Letters 12: 409-418). We have two questions. First, is there an
          > alternative to individual kill vials, given that we want insect-specific
          > pollen information? Working in the backcountry, it will be difficult to
          > handle the number of vials we'll likely need. Second, in removing pollen
          > from the insects' bodies, is the method described in Kearns and Inouye
          > 1993, i.e., rubbing a small cube of fuchsine stained gelatin over the
          > insect's body, still the best method? If so, how soon must this be done
          > after capture?
          >
          > If anyone has experience with mutualist networks in general, I'd love to
          > talk with you.
          >
          > Thanks for any advice!
          >
          > Diane
          > *******************************************
          > Diane L. Larson
          > Research Biologist
          > USGS Northern Prairie Wildlife Research Center
          > 1561 Lindig St.
          > St. Paul, MN 55108
          >
          > Voice 651-649-5041
          > FAX 651-649-5040
          > Email: dlarson@...
          >
          > ?It?s a poor sort of memory that only works backwards,? the Queen
          > remarked.
          > - Lewis Carroll, Through the Looking-Glass


          --
          Maria Stanko
          Graduate Program in Ecology and Evolution
          Dept. of Ecology, Evolution and Natural Resources
          Rutgers University
          Environmental & Natural Resources Building
          14 College Farm Road
          New Brunswick, New Jersey 08901
        • Doug Yanega
          OOH! OOH! OOH! (geeky kid in the back practically jumping out of his chair, waving his hand frantically for attention) I have the best technique EVER for
          Message 4 of 10 , Nov 10, 2009
          • 0 Attachment
            OOH! OOH! OOH!

            (geeky kid in the back practically jumping out of his chair, waving
            his hand frantically for attention)

            I have the best technique EVER for collecting pollen samples from
            bees. Really. And it doesn't even hurt the bees - you can let them go
            after the sample is taken (this was essential for me, as I used this
            technique for my thesis research, and I wanted to track how
            individual bees changed their pollen selection habits from day to
            day).

            It could hardly be simpler: it only requires a roll of Scotch Magic
            Transparent Tape (the kind that's clear with a matte finish), a
            pencil, and a box of microscope slides.

            You net the bee, remove her carefully, rub a small piece of tape
            against her belly until it picks up a smear of pollen, then you can
            let her go. As long as the piece of tape has pollen-free areas on
            either side of the pollen smear, you can simply stick it on the
            slide, and write directly on the tape with the pencil (indicating
            what bee it was, the time of day, and the locality/date). If you
            apply the pieces perpendicular to the long axis of the slide, you can
            fit up to 5 samples on a single slide. If a piece of tape sticks off
            the edge of the slide slightly, running a second slide along the edge
            slices off the excess flush with the edge.

            The ONLY trick to this (aside from not getting stung) is that one
            should not apply pressure directly to the portion of tape that has
            the pollen smear - it can crush and distort the grains. I wrote my
            data really small, and only at the ends of the pieces of tape where
            there was no pollen. I could gather dozens of samples a day this way,
            sometimes taking three or four samples from the same bee over the
            course of a day; the whole thing from net to slide takes about 30
            seconds once you're practiced at handling the bees without getting
            stung. I suppose one might try to position a bee within the net so
            the pollen grains can pass through the net mesh, to reduce the risk
            of stinging - but then one must have a very clean net bag so there is
            no chance of accidentally picking up residual pollen, and I doubt
            that's practical.

            This gives one a nice pollen sample to work with, with data written
            right there, and the pollen can either be examined directly by
            flipping the slide over, or - if one feels compelled to use
            traditional pollen-preparation techniques - small pieces of tape
            bearing pollen can be excised with an exacto-knife for processing
            (though this gives very small actual numbers of grains, so one's
            processing techniques have to be capable of working on tiny samples).

            I did this in 1983-6, and my slides are all still viable. The color
            of the pollen has faded somewhat, but other than that, they're pretty
            much unchanged. What I was able to do was wander around and take
            pollen samples directly from the anthers of the flowering plants in
            the vicinity, and then simply match the bee samples against the known
            pollen reference slides. I was able to determine that the bulk of
            pollen being collected was from flowering trees.

            That's it. No vials, no chemicals, no dead bees.

            If you use the technique and like it, just thank me in your
            acknowledgements. ;-)

            Peace,
            --

            Doug Yanega Dept. of Entomology Entomology Research Museum
            Univ. of California, Riverside, CA 92521-0314 skype: dyanega
            phone: (951) 827-4315 (standard disclaimer: opinions are mine, not UCR's)
            http://cache.ucr.edu/~heraty/yanega.html
            "There are some enterprises in which a careful disorderliness
            is the true method" - Herman Melville, Moby Dick, Chap. 82
          • Sam Droege
            Doug: Sounds like a good technique...You mentioned in another email some marking techniques for providing individual marks but I wonder if you have a quick
            Message 5 of 10 , Nov 11, 2009
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              Doug:

              Sounds like a good technique...You mentioned in another email some marking techniques for providing individual marks but I wonder if you have a quick marking technique so that you know that a  particular bee was sampled but isn't time consuming in a way that putting individually identifiable marks would be.  So, I am thinking...what about using a permanent marker pen and putting a dot on a forewing?.....any notions?

              sam

              From Field Work

              Not the mud slick,
              not the black weedy water
              full of alder cones and pock-marked leaves.

              Not the cow parsley in winter
              with its old whitened shins and wrists, its sibilance, its shaking.

              Not even the tart green shade of summer thick with butterflies
              and fungus plump as a leather saddle.

              No. But in a still corner,
              braced to its pebble-dashed wall,
              heavy, earth-drawn, all mouth and eye,

              the sunflower, dreaming umber.
              -Seamus Heaney




              From:Doug Yanega <dyanega@...>
              To:beemonitoring@yahoogroups.com
              Date:11/10/2009 01:04 PM
              Subject:Re: [beemonitoring] collecting pollen from bees
              Sent by:beemonitoring@yahoogroups.com





               

              OOH! OOH! OOH!

              (geeky kid in the back practically jumping out of his chair, waving
              his hand frantically for attention)

              I have the best technique EVER for collecting pollen samples from
              bees. Really. And it doesn't even hurt the bees - you can let them go
              after the sample is taken (this was essential for me, as I used this
              technique for my thesis research, and I wanted to track how
              individual bees changed their pollen selection habits from day to
              day).

              It could hardly be simpler: it only requires a roll of Scotch Magic
              Transparent Tape (the kind that's clear with a matte finish), a
              pencil, and a box of microscope slides.

              You net the bee, remove her carefully, rub a small piece of tape
              against her belly until it picks up a smear of pollen, then you can
              let her go. As long as the piece of tape has pollen-free areas on
              either side of the pollen smear, you can simply stick it on the
              slide, and write directly on the tape with the pencil (indicating
              what bee it was, the time of day, and the locality/date). If you
              apply the pieces perpendicular to the long axis of the slide, you can
              fit up to 5 samples on a single slide. If a piece of tape sticks off
              the edge of the slide slightly, running a second slide along the edge
              slices off the excess flush with the edge.

              The ONLY trick to this (aside from not getting stung) is that one
              should not apply pressure directly to the portion of tape that has
              the pollen smear - it can crush and distort the grains. I wrote my
              data really small, and only at the ends of the pieces of tape where
              there was no pollen. I could gather dozens of samples a day this way,
              sometimes taking three or four samples from the same bee over the
              course of a day; the whole thing from net to slide takes about 30
              seconds once you're practiced at handling the bees without getting
              stung. I suppose one might try to position a bee within the net so
              the pollen grains can pass through the net mesh, to reduce the risk
              of stinging - but then one must have a very clean net bag so there is
              no chance of accidentally picking up residual pollen, and I doubt
              that's practical.

              This gives one a nice pollen sample to work with, with data written
              right there, and the pollen can either be examined directly by
              flipping the slide over, or - if one feels compelled to use
              traditional pollen-preparation techniques - small pieces of tape
              bearing pollen can be excised with an exacto-knife for processing
              (though this gives very small actual numbers of grains, so one's
              processing techniques have to be capable of working on tiny samples).

              I did this in 1983-6, and my slides are all still viable. The color
              of the pollen has faded somewhat, but other than that, they're pretty
              much unchanged. What I was able to do was wander around and take
              pollen samples directly from the anthers of the flowering plants in
              the vicinity, and then simply match the bee samples against the known
              pollen reference slides. I was able to determine that the bulk of
              pollen being collected was from flowering trees.

              That's it. No vials, no chemicals, no dead bees.

              If you use the technique and like it, just thank me in your
              acknowledgements. ;-)

              Peace,
              --

              Doug Yanega Dept. of Entomology Entomology Research Museum
              Univ. of California, Riverside, CA 92521-0314 skype: dyanega
              phone: (951) 827-4315 (standard disclaimer: opinions are mine, not UCR's)

              http://cache.ucr.edu/~heraty/yanega.html
              "There are some enterprises in which a careful disorderliness
              is the true method" - Herman Melville, Moby Dick, Chap. 82



            • Anita M Collins, Ph.D.
              Sam, WE have successfully used the paint pens that are now available to mark bees when a quick mark,say just the color, is needed. Seems to stay on enough for
              Message 6 of 10 , Nov 11, 2009
              • 0 Attachment
                Sam,
                 
                WE have successfully used the paint pens that are now available to mark bees when a quick mark,say just the color, is needed.  Seems to stay on enough for some weeks that we can release newly emerged drones and catch them at sexual maturity. 
                 
                Anita
                If we knew what we were doing, it wouldn't be called research.
                -Albert Einstein





                Nov 11, 2009 06:20:51 AM, sdroege@... wrote:
                 


                Doug:

                Sounds like a good technique...You mentioned in another email some marking techniques for providing individual marks but I wonder if you have a quick marking technique so that you know that a  particular bee was sampled but isn't time consuming in a way that putting individually identifiable marks would be.  So, I am thinking...what about using a permanent marker pen and putting a dot on a forewing?.....any notions?

                sam

                From Field Work

                Not the mud slick,
                not the black weedy water
                full of alder cones and pock-marked leaves.

                Not the cow parsley in winter
                with its old whitened shins and wrists, its sibilance, its shaking.

                Not even the tart green shade of summer thick with butterflies
                and fungus plump as a leather saddle.

                No. But in a still corner,
                braced to its pebble-dashed wall,
                heavy, earth-drawn, all mouth and eye,

                the sunflower, dreaming umber.
                -Seamus Heaney




                From:Doug Yanega <dyanega@...>
                To:beemonitoring@yahoogroups.com
                Date:11/10/2009 01:04 PM
                Subject:Re: [beemonitoring] collecting pollen from bees
                Sent by:beemonitoring@yahoogroups.com





                 

                OOH! OOH! OOH!

                (geeky kid in the back practically jumping out of his chair, waving
                his hand frantically for attention)

                I have the best technique EVER for collecting pollen samples from
                bees. Really. And it doesn't even hurt the bees - you can let them go
                after the sample is taken (this was essential for me, as I used this
                technique for my thesis research, and I wanted to track how
                individual bees changed their pollen selection habits from day to
                day).

                It could hardly be simpler: it only requires a roll of Scotch Magic
                Transparent Tape (the kind that's clear with a matte finish), a
                pencil, and a box of microscope slides.

                You net the bee, remove her carefully, rub a small piece of tape
                against her belly until it picks up a smear of pollen, then you can
                let her go. As long as the piece of tape has pollen-free areas on
                either side of the pollen smear, you can simply stick it on the
                slide, and write directly on the tape with the pencil (indicating
                what bee it was, the time of day, and the locality/date). If you
                apply the pieces perpendicular to the long axis of the slide, you can
                fit up to 5 samples on a single slide. If a piece of tape sticks off
                the edge of the slide slightly, running a second slide along the edge
                slices off the excess flush with the edge.

                The ONLY trick to this (aside from not getting stung) is that one
                should not apply pressure directly to the portion of tape that has
                the pollen smear - it can crush and distort the grains. I wrote my
                data really small, and only at the ends of the pieces of tape where
                there was no pollen. I could gather dozens of samples a day this way,
                sometimes taking three or four samples from the same bee over the
                course of a day; the whole thing from net to slide takes about 30
                seconds once you're practiced at handling the bees without getting
                stung. I suppose one might try to position a bee within the net so
                the pollen grains can pass through the net mesh, to reduce the risk
                of stinging - but then one must have a very clean net bag so there is
                no chance of accidentally picking up residual pollen, and I doubt
                that's practical.

                This gives one a nice pollen sample to work with, with data written
                right there, and the pollen can either be examined directly by
                flipping the slide over, or - if one feels compelled to use
                traditional pollen-preparation techniques - small pieces of tape
                bearing pollen can be excised with an exacto-knife for processing
                (though this gives very small actual numbers of grains, so one's
                processing techniques have to be capable of working on tiny samples).

                I did this in 1983-6, and my slides are all still viable. The color
                of the pollen has faded somewhat, but other than that, they're pretty
                much unchanged. What I was able to do was wander around and take
                pollen samples directly from the anthers of the flowering plants in
                the vicinity, and then simply match the bee samples against the known
                pollen reference slides. I was able to determine that the bulk of
                pollen being collected was from flowering trees.

                That's it. No vials, no chemicals, no dead bees.

                If you use the technique and like it, just thank me in your
                acknowledgements. ;-)

                Peace,
                --

                Doug Yanega Dept. of Entomology Entomology Research Museum
                Univ. of California, Riverside, CA 92521-0314 skype: dyanega
                phone: (951) 827-4315 (standard disclaimer: opinions are mine, not UCR's)
                http://cache.ucr.edu/~heraty/yanega.html
                "There are some enterprises in which a careful disorderliness
                is the true method" - Herman Melville, Moby Dick, Chap. 82



              • Jack Neff
                Sam:  I also have marked many bees of varying sizes with fast drying enamels and paint pens. I found it easier said than done. Being a clutz, I regularly
                Message 7 of 10 , Nov 11, 2009
                • 0 Attachment
                  Sam:  I also have marked many bees of varying sizes with fast drying enamels and paint pens. I found it easier said than done. Being a clutz, I regularly painted the wing bases with the results Doug mentioned.  Chilling the bees in a cooler slows them down and makes them easier to mark.  I also had a problem with fading with paint pens so that the lighter colors became indistinguishable after a few weeks.  The "Techniques for Pollination Biologists" book by Kearns and Inouye has 11 pages on marking insects, although much of that ground has already been by other discussants.

                  best

                  Jack

                  John L. Neff
                  Central Texas Melittological Institute
                  7307 Running Rope
                  Austin,TX 78731 USA
                  512-345-7219

                  --- On Wed, 11/11/09, Anita M Collins, Ph.D. <frozenbeedoc@...> wrote:

                  From: Anita M Collins, Ph.D. <frozenbeedoc@...>
                  Subject: Re: Re: [beemonitoring] collecting pollen from bees
                  To: sdroege@...
                  Cc: beemonitoring@yahoogroups.com
                  Date: Wednesday, November 11, 2009, 9:52 AM

                   

                  Sam,
                   
                  WE have successfully used the paint pens that are now available to mark bees when a quick mark,say just the color, is needed.  Seems to stay on enough for some weeks that we can release newly emerged drones and catch them at sexual maturity. 
                   
                  Anita
                  If we knew what we were doing, it wouldn't be called research.
                  -Albert Einstein





                  Nov 11, 2009 06:20:51 AM, sdroege@usgs. gov wrote:
                   


                  Doug:

                  Sounds like a good technique... You mentioned in another email some marking techniques for providing individual marks but I wonder if you have a quick marking technique so that you know that a  particular bee was sampled but isn't time consuming in a way that putting individually identifiable marks would be.  So, I am thinking...what about using a permanent marker pen and putting a dot on a forewing?... ..any notions?

                  sam

                  From Field Work

                  Not the mud slick,
                  not the black weedy water
                  full of alder cones and pock-marked leaves.

                  Not the cow parsley in winter
                  with its old whitened shins and wrists, its sibilance, its shaking.

                  Not even the tart green shade of summer thick with butterflies
                  and fungus plump as a leather saddle.

                  No. But in a still corner,
                  braced to its pebble-dashed wall,
                  heavy, earth-drawn, all mouth and eye,

                  the sunflower, dreaming umber.
                  -Seamus Heaney




                  From:Doug Yanega <dyanega@ucr. edu>
                  To:beemonitoring@ yahoogroups. com
                  Date:11/10/2009 01:04 PM
                  Subject:Re: [beemonitoring] collecting pollen from bees
                  Sent by:beemonitoring@ yahoogroups. com





                   

                  OOH! OOH! OOH!

                  (geeky kid in the back practically jumping out of his chair, waving
                  his hand frantically for attention)

                  I have the best technique EVER for collecting pollen samples from
                  bees. Really. And it doesn't even hurt the bees - you can let them go
                  after the sample is taken (this was essential for me, as I used this
                  technique for my thesis research, and I wanted to track how
                  individual bees changed their pollen selection habits from day to
                  day).

                  It could hardly be simpler: it only requires a roll of Scotch Magic
                  Transparent Tape (the kind that's clear with a matte finish), a
                  pencil, and a box of microscope slides.

                  You net the bee, remove her carefully, rub a small piece of tape
                  against her belly until it picks up a smear of pollen, then you can
                  let her go. As long as the piece of tape has pollen-free areas on
                  either side of the pollen smear, you can simply stick it on the
                  slide, and write directly on the tape with the pencil (indicating
                  what bee it was, the time of day, and the locality/date) . If you
                  apply the pieces perpendicular to the long axis of the slide, you can
                  fit up to 5 samples on a single slide. If a piece of tape sticks off
                  the edge of the slide slightly, running a second slide along the edge
                  slices off the excess flush with the edge.

                  The ONLY trick to this (aside from not getting stung) is that one
                  should not apply pressure directly to the portion of tape that has
                  the pollen smear - it can crush and distort the grains. I wrote my
                  data really small, and only at the ends of the pieces of tape where
                  there was no pollen. I could gather dozens of samples a day this way,
                  sometimes taking three or four samples from the same bee over the
                  course of a day; the whole thing from net to slide takes about 30
                  seconds once you're practiced at handling the bees without getting
                  stung. I suppose one might try to position a bee within the net so
                  the pollen grains can pass through the net mesh, to reduce the risk
                  of stinging - but then one must have a very clean net bag so there is
                  no chance of accidentally picking up residual pollen, and I doubt
                  that's practical.

                  This gives one a nice pollen sample to work with, with data written
                  right there, and the pollen can either be examined directly by
                  flipping the slide over, or - if one feels compelled to use
                  traditional pollen-preparation techniques - small pieces of tape
                  bearing pollen can be excised with an exacto-knife for processing
                  (though this gives very small actual numbers of grains, so one's
                  processing techniques have to be capable of working on tiny samples).

                  I did this in 1983-6, and my slides are all still viable. The color
                  of the pollen has faded somewhat, but other than that, they're pretty
                  much unchanged. What I was able to do was wander around and take
                  pollen samples directly from the anthers of the flowering plants in
                  the vicinity, and then simply match the bee samples against the known
                  pollen reference slides. I was able to determine that the bulk of
                  pollen being collected was from flowering trees.

                  That's it. No vials, no chemicals, no dead bees.

                  If you use the technique and like it, just thank me in your
                  acknowledgements. ;-)

                  Peace,
                  --

                  Doug Yanega Dept. of Entomology Entomology Research Museum
                  Univ. of California, Riverside, CA 92521-0314 skype: dyanega
                  phone: (951) 827-4315 (standard disclaimer: opinions are mine, not UCR's)
                  http://cache.ucr.edu/~heraty/ yanega.html
                  "There are some enterprises in which a careful disorderliness
                  is the true method" - Herman Melville, Moby Dick, Chap. 82




                • Sam Droege
                  Thanks Jack: I was thinking that Doug s technique sounds useful, but you would need to know if you had sampled that bee already (at least that day) or not but
                  Message 8 of 10 , Nov 11, 2009
                  • 0 Attachment

                    Thanks Jack:

                    I was thinking that Doug's technique sounds useful, but you would need to know if you had sampled that bee already (at least that day) or not but it would slow things down too much if there was a lot of futzing around.  Additionally, have a dead body means that I can verify (and others after me) all identifications.  In general I think that ID'ing bees in the field on the wing or even in the hand is problematic even when its someone with lots of experience...so, am naturally leaning towards kill jars.

                    sam

                    Sam Droege  sdroege@...                      
                    w 301-497-5840 h 301-390-7759 fax 301-497-5624
                    USGS Patuxent Wildlife Research Center
                    BARC-EAST, BLDG 308, RM 124 10300 Balt. Ave., Beltsville, MD  20705
                    Http://www.pwrc.usgs.gov

                    More black than ash-buds in the front of March.
                      --Al Tennyson






                    From:Jack Neff <jlnatctmi@...>
                    To:sdroege@..., " Ph.D.Anita M Collins" <frozenbeedoc@...>
                    Cc:beemonitoring@yahoogroups.com
                    Date:11/11/2009 11:11 AM
                    Subject:Re: Re: [beemonitoring] collecting pollen from bees
                    Sent by:beemonitoring@yahoogroups.com





                     

                    Sam:  I also have marked many bees of varying sizes with fast drying enamels and paint pens. I found it easier said than done. Being a clutz, I regularly painted the wing bases with the results Doug mentioned.  Chilling the bees in a cooler slows them down and makes them easier to mark.  I also had a problem with fading with paint pens so that the lighter colors became indistinguishable after a few weeks.  The "Techniques for Pollination Biologists" book by Kearns and Inouye has 11 pages on marking insects, although much of that ground has already been by other discussants.

                    best

                    Jack

                    John L. Neff
                    Central Texas Melittological Institute
                    7307 Running Rope
                    Austin,TX 78731 USA
                    512-345-7219

                    --- On Wed, 11/11/09, Anita M Collins, Ph.D. <frozenbeedoc@...> wrote:


                    From: Anita M Collins, Ph.D. <frozenbeedoc@...>
                    Subject: Re: Re: [beemonitoring] collecting pollen from bees
                    To: sdroege@...
                    Cc: beemonitoring@yahoogroups.com
                    Date: Wednesday, November 11, 2009, 9:52 AM

                     

                    Sam,
                     
                    WE have successfully used the paint pens that are now available to mark bees when a quick mark,say just the color, is needed.  Seems to stay on enough for some weeks that we can release newly emerged drones and catch them at sexual maturity.  
                     
                    Anita
                    If we knew what we were doing, it wouldn't be called research.
                    -Albert Einstein





                    Nov 11, 2009 06:20:51 AM,
                    sdroege@usgs. gov wrote:
                     


                    Doug:


                    Sounds like a good technique... You mentioned in another email some marking techniques for providing individual marks but I wonder if you have a quick marking technique so that you know that a  particular bee was sampled but isn't time consuming in a way that putting individually identifiable marks would be.  So, I am thinking...what about using a permanent marker pen and putting a dot on a forewing?... ..any notions?

                    sam

                    From Field Work

                    Not the mud slick,
                    not the black weedy water
                    full of alder cones and pock-marked leaves.

                    Not the cow parsley in winter
                    with its old whitened shins and wrists, its sibilance, its shaking.

                    Not even the tart green shade of summer thick with butterflies
                    and fungus plump as a leather saddle.

                    No. But in a still corner,
                    braced to its pebble-dashed wall,
                    heavy, earth-drawn, all mouth and eye,

                    the sunflower, dreaming umber.
                    -Seamus Heaney




                    From: Doug Yanega <dyanega@ucr. edu>
                    To: beemonitoring@ yahoogroups. com
                    Date: 11/10/2009 01:04 PM
                    Subject: Re: [beemonitoring] collecting pollen from bees
                    Sent by: beemonitoring@ yahoogroups. com





                     

                    OOH! OOH! OOH!

                    (geeky kid in the back practically jumping out of his chair, waving
                    his hand frantically for attention)

                    I have the best technique EVER for collecting pollen samples from
                    bees. Really. And it doesn't even hurt the bees - you can let them go
                    after the sample is taken (this was essential for me, as I used this
                    technique for my thesis research, and I wanted to track how
                    individual bees changed their pollen selection habits from day to
                    day).

                    It could hardly be simpler: it only requires a roll of Scotch Magic
                    Transparent Tape (the kind that's clear with a matte finish), a
                    pencil, and a box of microscope slides.

                    You net the bee, remove her carefully, rub a small piece of tape
                    against her belly until it picks up a smear of pollen, then you can
                    let her go. As long as the piece of tape has pollen-free areas on
                    either side of the pollen smear, you can simply stick it on the
                    slide, and write directly on the tape with the pencil (indicating
                    what bee it was, the time of day, and the locality/date) . If you
                    apply the pieces perpendicular to the long axis of the slide, you can
                    fit up to 5 samples on a single slide. If a piece of tape sticks off
                    the edge of the slide slightly, running a second slide along the edge
                    slices off the excess flush with the edge.

                    The ONLY trick to this (aside from not getting stung) is that one
                    should not apply pressure directly to the portion of tape that has
                    the pollen smear - it can crush and distort the grains. I wrote my
                    data really small, and only at the ends of the pieces of tape where
                    there was no pollen. I could gather dozens of samples a day this way,
                    sometimes taking three or four samples from the same bee over the
                    course of a day; the whole thing from net to slide takes about 30
                    seconds once you're practiced at handling the bees without getting
                    stung. I suppose one might try to position a bee within the net so
                    the pollen grains can pass through the net mesh, to reduce the risk
                    of stinging - but then one must have a very clean net bag so there is
                    no chance of accidentally picking up residual pollen, and I doubt
                    that's practical.

                    This gives one a nice pollen sample to work with, with data written
                    right there, and the pollen can either be examined directly by
                    flipping the slide over, or - if one feels compelled to use
                    traditional pollen-preparation techniques - small pieces of tape
                    bearing pollen can be excised with an exacto-knife for processing
                    (though this gives very small actual numbers of grains, so one's
                    processing techniques have to be capable of working on tiny samples).

                    I did this in 1983-6, and my slides are all still viable. The color
                    of the pollen has faded somewhat, but other than that, they're pretty
                    much unchanged. What I was able to do was wander around and take
                    pollen samples directly from the anthers of the flowering plants in
                    the vicinity, and then simply match the bee samples against the known
                    pollen reference slides. I was able to determine that the bulk of
                    pollen being collected was from flowering trees.

                    That's it. No vials, no chemicals, no dead bees.

                    If you use the technique and like it, just thank me in your
                    acknowledgements. ;-)

                    Peace,
                    --

                    Doug Yanega Dept. of Entomology Entomology Research Museum
                    Univ. of California, Riverside, CA 92521-0314 skype: dyanega
                    phone: (951) 827-4315 (standard disclaimer: opinions are mine, not UCR's)

                    http://cache.ucr.edu/~heraty/ yanega.html
                    "There are some enterprises in which a careful disorderliness
                    is the true method" - Herman Melville, Moby Dick, Chap. 82




                  • Laura Burkle
                    Sam and all, I don t have too much more to add that others haven t already mentioned. But I will put a plug in for using individual vials to collect bees from
                    Message 9 of 10 , Nov 12, 2009
                    • 0 Attachment
                      Sam and all,

                      I don't have too much more to add that others haven't already
                      mentioned. But I will put a plug in for using individual vials to
                      collect bees from which you'd like to get pollen from later for network
                      analysis, etc. We use small 3.5 dram vials from Thornton Plastics or
                      standard 1.7mL microtubes, depending on the size of the bee. One can
                      carry a lot of these vials in the field, and you won't risk pollen
                      "contamination" between bees like you might in kill jars. We freeze the
                      bees in a cooler while we're in the field, and later process the dead
                      bees for pollen, rubbing the bee with a fuchsin-gelatin cube or washing
                      the bee in ethanol and staining the pollen with the fuchsin dye once the
                      ethanol has evaporated from the slide.

                      As for marking, we use different colors of Deco paint pens (extra
                      fine or fine tip) on the thorax. Paint pen dots seem to last at least
                      three weeks. We use a bee-squeezer (film canister or other vial with
                      the bottom cut out, mesh covering over the top, and a foam plunger to
                      push the bee against the mesh) to position the bee and mark through the
                      mesh.


                      Best ---

                      Laura


                      Postdoctoral Research Associate

                      Washington University in St. Louis
                      (314) 935-9445
                      http://biology4.wustl.edu/burkle/
                    • Diane L Larson
                      Thanks to everyone for all the great information on pollen collection! Diane ******************************************* Diane L. Larson Research Biologist
                      Message 10 of 10 , Nov 12, 2009
                      • 0 Attachment

                        Thanks to everyone for all the great information on pollen collection!

                        Diane
                        *******************************************
                        Diane L. Larson
                        Research Biologist
                        USGS Northern Prairie Wildlife Research Center
                        1561 Lindig St.
                        St. Paul, MN  55108

                        Voice 651-649-5041
                        FAX 651-649-5040
                        Email: dlarson@...

                        “It’s a poor sort of memory that only works backwards,” the Queen remarked.
                        - Lewis Carroll, Through the Looking-Glass



                        From:Laura Burkle <laura.a.burkle.adv08@...>
                        To:beemonitoring@yahoogroups.com
                        Date:11/12/2009 09:01 AM
                        Subject:[beemonitoring] Re: collecting pollen from bees
                        Sent by:beemonitoring@yahoogroups.com





                         

                        Sam and all,

                        I don't have too much more to add that others haven't already
                        mentioned. But I will put a plug in for using individual vials to
                        collect bees from which you'd like to get pollen from later for network
                        analysis, etc. We use small 3.5 dram vials from Thornton Plastics or
                        standard 1.7mL microtubes, depending on the size of the bee. One can
                        carry a lot of these vials in the field, and you won't risk pollen
                        "contamination" between bees like you might in kill jars. We freeze the
                        bees in a cooler while we're in the field, and later process the dead
                        bees for pollen, rubbing the bee with a fuchsin-gelatin cube or washing
                        the bee in ethanol and staining the pollen with the fuchsin dye once the
                        ethanol has evaporated from the slide.

                        As for marking, we use different colors of Deco paint pens (extra
                        fine or fine tip) on the thorax. Paint pen dots seem to last at least
                        three weeks. We use a bee-squeezer (film canister or other vial with
                        the bottom cut out, mesh covering over the top, and a foam plunger to
                        push the bee against the mesh) to position the bee and mark through the
                        mesh.

                        Best ---

                        Laura

                        Postdoctoral Research Associate

                        Washington University in St. Louis
                        (314) 935-9445

                        http://biology4.wustl.edu/burkle/



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