Cryo-Confocal Imaging
the larger tissue slice to enable correlation with subsequent EM imaging. As we describe here, the integration of cryo-CLSM of tissue samples into the CLEM workflow opened a door for bet- ter understanding of biological phenomena that require rapid temporal resolution to analyze. Te introduction of geneti- cally encoded EM markers in the tissue will be the next step to eliminate the immuno-EM labeling process from the current workflow.
Conclusions In this paper, we integrate cryo-confocal microscopy of
frozen brain slices into a CLEM workflow to assist the iden- tification of target cellular features between 2-photon LM and EM. We show that it is possible to image frozen tissue under cryogenic conditions and demonstrate the viability of Airyscan detection for high-SNR imaging.
Acknowledgements Tis work was supported by the Max Planck Florida
Institute for Neuroscience. We would like to thank Carl Zeiss Microscopy, LLC and Linkam Scientific Instruments for providing the Cryo-Correlative Microscopy Stage and with developing the prototype of the carrier adaptor; we also thank them for their kind technical support through the Research Partnership between Carl Zeiss Microscopy, LLC and MPFI.
References [1] C Humpel, Neuroscience 305 (2015)
https://doi.org/10.1016/ j.neuroscience.2015.07.086.
[2] K Harris and SB Kater, Ann Rev Neurosci 17 (1994) https://
doi.org/10.1146/annurev.ne.17.030194.002013.
[3] R Lamprecht and J LeDoux, Nat Rev Neurosci 5 (2004)
https://doi.org/10.1038/nrn1301.
[4] HB Kwon and BL Sabatini, Nature 474 (2011) https://doi .org/10.1038/nature09986.
[5] J Noguchi et al., J Physiol 589 (2011)
https://doi.org/10.1113/ jphysiol.2011.207100.
[6] Y Sun et al., “Correlative ultrastructural analysis of func- tionally modulated synapses using automated tape-collect- ing ultramicrotome and SEM array tomography,” Chapter 7 in: Volume microscopy, I Wacker, et al., eds. Humana, New York, (2020)
https://doi.org/10.1007/978-1-0716-0691-9_7.
[7] Y Sun et al., J Neurosci (July 15, 2021) https://doi .org/10.1523/jneurosci.1964-20.2021.
[8] D Studer et al., Histochem Cell Biol 130 (2008) https://doi .org/10.1007/s00418-008-0500-1.
[9] R Kaufmann et al., Curr Opin Chem Biol 20 (2014) https://
doi.org/10.1016/j.cbpa.2014.05.007.
[10] GH Wu et al., Structure 28 (2020)
https://doi.org/10.1016/ j.str.2020.07.017.
[11] S Klein et al., Nat Commun Biol 4 (2021) https://doi .org/10.1038/s42003-020-01567-z.
[12] J Kuba et al., J Microsc 281 (2021)
https://doi.org/10.1111/ jmi.12939.
[13] J Schindelin et al., Nat Methods 9 (2012) https://doi .org/10.1038/nmeth.2019.
Show Your FIB System Some Love High quality extractors, suppressors and aperture strips from Oregon Physics
All parts are made to exacting quality standards for guaranteed performance in your system.
+1 503 601 0041
info@oregon-physics.com
www.oregon-physics.com
2021 September •
www.microscopy-today.com
Ad_Oregon-Physics-Parts-v012721.indd 1 1/27/21 4:52 PM 39
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80
