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Re: [beemonitoring] collecting pollen from bees

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  • 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 1 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 2 of 10 , Nov 10, 2009
      • 0 Attachment
        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 3 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 4 of 10 , Nov 11, 2009
          • 0 Attachment

            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 5 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 6 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 7 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 8 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 9 of 10 , Nov 12, 2009
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                      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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