3.3 Histological
Tissue Staining for
Laser Capture
Microdissection
Transport slides (in slide boxes) on dry ice. Use forceps to move
slides between staining containers, wear gloves, and clean these
regularly with RNaseZap. For examples of histological stainings
seeFig. 1.- Before initiating the staining, switch on the LCM system
(microscope, laser, computer, start LMD software) and turn
on the UV light and fan in the LCM chamber if available.
Fig. 1Examples of HistoGene and cresyl violet stainings of mouse and human spinal cords at different
magnifications. (a–d, i–l) Note that the HistoGene staining in general gives a better contrast. (e–h, m–p) Small
nonneuronal cells are more visible after staining with cresyl violet. (a, e, i, m) Images taken at 5
magnification provide a good overview of the tissue morphology for reference. (b, f, j, n) Images taken
prior to microdissection at 20magnification are a helpful reference to identify cells of interest. (c, g, k, o)
Note the slightly decreased contrast at 40magnification with less distinct nuclei of the cells. To avoid
contamination with surrounding cells, keep cutting outlines close to the cells. (d, h, l, p) Tissue after isolation
of cells. Scale bar inm(applicable toa, e, i, m)¼ 400 μm, inn(applicable tob, f, j, n)¼ 100 μm and in
p(applicable toc, d, g, h, k, l, o, p)¼ 50 μm)
Spatial Transcriptomic Profiling Using LCM-seq 101