Collection of Epithelial Cells from Rodent Mammary Gland Via Laser Capture Microdissection Yielding High-Quality RNA Suitable for Microarray Analysis

Table 1 RNA yield and LCM cell estimate

Tissue	Experion^a(ng/μL)	NanoDrop (ng/μL)	Total RNA (ng)	Laser fires per cap	Cell Count per cap^b	Cell Count per sample^c
MA (n = 27)	7.9 ± 0.8 (2.1, 16.7)	8.6 ± 0.8 (2.3, 17.8)	86	3,277.9 ± 141.0 (778.0; 8,212.0)	6,202.2 ± 223.2 (1,182.6; 12,511.5)	24,808.7 ± 1,270.2
MG (n = 27)	4.5 ± 0.4 (1.8, 10.4)	4.9 ± 0.5 (1.8, 14.5)	49	785.6 33.0 (61.0; 2,190.0)	994.2 ± 42.5 (69.5; 2,678.2)	8,192.2 ± 728.3

^aValues are means ± SEM (min, max). Good correlation was observed between Experion and NanoDrop results obtained from both MA (r = 0.88, p < 0.01) and MG (r = 0.83, p < 0.01). Cell estimates were calculated based on the formula developed by Espina et al. [10]. Spot sizes ranged from 20 to 40 μm for MA and 10 to 15 μm for MG. A typical cell diameter of 7 μm was used for all rat mammary epithelium, percent overlap was kept constant at 40%, and 95% efficiency was assumed
^bBecause of limitations in the amount of time that could be spent collecting cells on each cap without RNA degradation and the dispensed nature of cell distribution within MG, there were fewer cells per MG cap and more variability in cell count per cap
^cA total of four caps were dissected each MA and eight caps from each MG specimen

ISSN: 1480-9222