scholarly journals Immunohistochemical staining, laser capture microdissection, and filter-aided sample preparation-assisted proteomic analysis of target cell populations within tissue samples

2013 ◽  
Vol 34 (11) ◽  
pp. 1627-1636 ◽  
Author(s):  
Jintang He ◽  
Jianhui Zhu ◽  
Yashu Liu ◽  
Jing Wu ◽  
Song Nie ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Target Cell ◽  
Laser Capture Microdissection ◽  
Proteomic Analysis ◽  
Immunohistochemical Staining ◽  
Cell Populations ◽  
Tissue Samples ◽  
Laser Capture

Proteomic Analysis of Frozen Tissue Samples Using Laser Capture Microdissection

2013 ◽  
pp. 71-83 ◽  
Author(s):  
Sumana Mukherjee ◽  
Jaime Rodriguez-Canales ◽  
Jeffrey Hanson ◽  
Michael R. Emmert-Buck ◽  
Michael A. Tangrea ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Proteomic Analysis ◽  
Tissue Samples ◽  
Frozen Tissue ◽  
Laser Capture

scholarly journals An optimised protocol for isolation of RNA through laser capture microdissection of leaf material

2019 ◽  
Author(s):  
Lei Hua ◽  
Julian M Hibberd
Keyword(s):  
Sample Preparation ◽  
Laser Capture Microdissection ◽  
Leaf Tissue ◽  
Cell Types ◽  
Rna Degradation ◽  
Mesophyll Cells ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Aqueous Conditions ◽  
Laser Capture

AbstractLaser Capture Microdissection is a powerful tool that allows thin slices of specific cells types to be separated from one another. However, the most commonly used protocol, which involves embedding tissue in paraffin wax, results in severely degraded RNA. Yields from low abundance cell types of leaves are particularly compromised. We reasoned that the relatively high temperature used for sample embedding, and aqueous conditions associated with sample preparation prior to microdissection contribute to RNA degradation. Here we describe an optimized procedure to limit RNA degradation that is based on the use of low melting point wax as well as modifications to sample preparation prior to dissection, and isolation of paradermal, rather than transverse sections. Using this approach high quality RNA suitable for down-stream applications such as quantitative reverse transcriptase polymerase chain reactions or RNA-sequencing is recovered from microdissected bundle sheath strands and mesophyll cells of leaf tissue.

scholarly journals Optimizing Laser Capture Microdissection Protocol for Isolating Zone-Specific Cell Populations from Mandibular Condylar Cartilage

2019 ◽  
Vol 2019 ◽  
pp. 1-13
Author(s):  
Aisha M. Basudan ◽  
Yanqi Yang
Keyword(s):  
Laser Capture Microdissection ◽  
Genetic Data ◽  
Male Rats ◽  
Cell Populations ◽  
Specific Cell ◽  
Condylar Cartilage ◽  
Rna Integrity ◽  
Tissue Sections ◽  
Mandibular Condylar Cartilage ◽  
Laser Capture

Mandibular condylar cartilage (MCC) is a multizonal heterogeneous fibrocartilage consisting of fibrous (FZ), proliferative (PZ), mature (MZ), and hypertrophic (HZ) zones. Gross sampling of the whole tissue may conceal some important information and compromise the validity of the molecular analysis. Laser capture microdissection (LCM) technology allows isolating zonal (homogenous) cell populations and consequently generating more accurate molecular and genetic data, but the challenges during tissue preparation and microdissection procedures are to obtain acceptable tissue section morphology that allows histological identification of the desirable cell type and to minimize RNA degradation. Therefore, our aim is to optimize an LCM protocol for isolating four homogenous zone-specific cell populations from their respective MCC zones while preserving the quality of RNA recovered. MCC and FCC (femoral condylar cartilage) specimens were harvested from 5-week-old Sprague–Dawley male rats. Formalin-fixed and frozen unfixed tissue sections were prepared and compared histologically. Additional specimens were microdissected to prepare LCM samples from FCC and each MCC zone individually. Then, to evaluate LCM-RNA integrity, 3′/m ratios of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (β-Actin) using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were calculated. Both fixed and unfixed tissue sections allowed reliable identification of MCC zones. The improved morphology of the frozen sections of our protocol has extended the range of cell types to be isolated. Under the empirically set LCM parameters, four homogeneous cell populations were efficiently isolated from their respective zones. The 3′/m ratio means of GAPDH and β-Actin ranged between 1.11–1.56 and 1.41–2.12, respectively. These values are in line with the reported quality control requirements. The present study shows that the optimized LCM protocol could allow isolation of four homogenous zone-specific cell populations from MCC, meanwhile preserving RNA integrity to meet the high quality requirements for subsequent molecular analyses. Thereby, accurate molecular and genetic data could be generated.

LASER CAPTURE MICRODISSECTION AND PROTEOMIC ANALYSIS — PRELIMINARY FINDINGS

1999 ◽  
Vol 27 (3) ◽  
pp. A67-A67
Author(s):  
R E Banks ◽  
M J Dunn ◽  
M A Forbes ◽  
A Stanley ◽  
D Pappin ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Proteomic Analysis ◽  
Laser Capture
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