- Allows identification of target cells without compromising RNA quality or LCM efficiency
- Maintenance of RNA integrity ensures exceptional performance in downstream applications
- Includes a stain that clearly shows cell distribution – ideal for cases where tissue morphology has been disrupted
- Use with Ambion’s RNAqueous®-Micro and MessageAmp II Kits for superior gene expression analyses
Laser Capture Microdissection (LCM) is a method for obtaining pure populations of cells from heterogeneous samples. LCM permits the selection and capture of cells, cell aggregates, and discrete morphological structures from thin tissue sections. Captured cells can then be used for nucleic acid studies, including SNP analysis, endpoint and real-time RT-PCR, and mRNA expression profiling. It is often necessary to stain tissue sections so that discrete structures within the tissue can be discerned. To isolate RNA from LCM samples, it is important to use a procedure that minimizes RNA degradation during staining.
To circumvent the RNA degradation that occurs with traditional staining protocols, Ambion’s LCM specialists developed a novel staining procedure that avoids exposing the tissue sections to pure water at any step. (See Figure 1)
The procedure has been incorporated into a kit that provides superior staining of frozen sections to facilitate identification of target cells for LCM, while preserving RNA quality.
The LCM Staining Kit provides enough reagents for the processing of 80 slides. Two different stains are provided with the kit: Cresyl Violet and Acridine Orange.
Provide Superior Staining While Maintaining RNA Integrity
Tissues prepared for LCM using the LCM Staining Kit yield high quality RNA suitable for qRT-PCR and microarrays. (See Figure 2)
Obtain High Quality RNA from Many Tissue Types
LCM Specialists at Ambion have tested the LCM Staining kit with a variety of tissue types including mouse brain, liver, spleen, and kidney. Human tissues include ovarian tumor, uterine tumor, lymphoma, small intestine, colon and breast. (See Figure 3)
Easy Identification of Target Cells
The Cresyl Violet stain allows the user to visualize variations in cell morphology which can be useful in identifying malignant cells. In addition, using the Acridine Orange stain offers an advantage when microdissecting single cells, where the visualization of the tissue captured in the cap is not very clear due to the melting of the thermoplastic film. With Acridine Orange, isolated cells will fluoresce on the cap after microdissection, making it easy to assess whether the tissue/cell was effectively microdissected and retrieved. (See Figure 4)
LCM Staining Kit Components:
Acridine Orange stain
Cresyl Violet stain
Slide chambers
Dehydration beads
RNaseZap®
500 ml bottles
Barrier pen
Nuclease-free water
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