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Laser-capture microdissection

PL Peipei Lu
JF Joseph Foley
CZ Chunfang Zhu
KM Katherine McNamara
KS Korsuk Sirinukunwattana
SV Sujay Vennam
SV Sushama Varma
HF Hamid Fehri
AS Arunima Srivastava
SZ Shirley Zhu
JR Jens Rittscher
PM Parag Mallick
CC Christina Curtis
RW Robert West
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Consecutive sections of the FFPE block were cut on a microtome at 7-μm thickness and mounted on glass slides with polyethylene naphthalate membranes (Thermo Fisher Scientific LCM0522). Slides were immersed 20 s each in xylenes×3, 100% ethanol×3, 95% ethanol×2, 70% ethanol×2, water, hematoxylin (Dako S3309), water, bluing reagent (Thermo Fisher Scientific 7301), water, 70% ethanol×2, 95% ethanol×2, and 100% ethanol×3, xylenes×3. Slides were dissected immediately after staining. Cells were dissected on an ArcturusXT LCM System using both the ultraviolet (UV) laser to cut out each sample and the infrared laser to adhere it to a CapSure HS LCM Cap (Thermo Fisher Scientific LCM0215). Roughly 500 cells were captured by area, according to density estimates by cell counting on small areas. After LCM, the cap was sealed in a 0.5-mL tube (Thermo Fisher Scientific N8010611) and subjected to Smart-3SEQ or DNA library preparation.

Copyright and License information: The Author(s) ©2021
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