Tissue Preparation for Laser Capture Microdissection

All samples for LCM were cut with a microtome to 10 µm slices (Leica, RM2055) and were mounted on glass slides (SuperFrost Ultra Plus, Menzel Gläser, Thermofisher Scientific [33 (link)]) with a drop of DNAse/RNAse-free water. Next, samples were incubated in a fume hood at 56 °C for 1 h to increase adherence to slides. Mounted slices were hematoxylin stained according to the standard protocol in a set of alcohol solutions, xylene, and stain.

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Grzywa T.M., Koppolu A.A., Paskal W., Klicka K., Rydzanicz M., Wejman J., Płoski R, & Włodarski P.K. (2021). Higher Mutation Burden in High Proliferation Compartments of Heterogeneous Melanoma Tumors. International Journal of Molecular Sciences, 22(8), 3886.

Publication 2021

RM2055

Alcohol Dnase Hematoxylin Microtome Rnase Stain Xylene

Corresponding Organization : Medical University of Warsaw

Other organizations : Postgraduate School of Molecular Medicine

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Variable analysis

independent variables

Microtome type (Leica, RM2055)

dependent variables

Adherence of mounted slices to glass slides

control variables

Slice thickness (10 µm)
Slide type (SuperFrost Ultra Plus, Menzel Gläser, Thermofisher Scientific)
Incubation time (1 h)
Incubation temperature (56 °C)
Staining protocol (hematoxylin stained according to the standard protocol in a set of alcohol solutions, xylene, and stain)

controls

No positive or negative controls were explicitly mentioned.

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