Tissue Staining

The Arcturus® HistoGene® LCM Frozen Section Staining Kit comes complete with all the reagents and supplies needed for preparing frozen tissue sections for laser capture microdissection (LCM). Designed as a fast-penetrating stain, this kit provides excellent contrast while preserving nucleic acid integrity and retaining low-abundance mRNAs.

Each kit contains:

• HistoGene® Staining Solution
• 100%, 95%, 75% ethanol
• Xylene
• Disposable slide staining jar
• HistoGene® LCM Slides
• Validated protocol

Retain Low-Abundance mRNA
RT-PCR analysis of specific genes from cells captured from tissues processed using the HistoGene® kit shows retention of both low-abundance mRNA and higher-abundance species (Figure 1). The mRNA profile of HistoGene® Kit samples appears free of degradation.

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Figure 1. Detection of low-abundance mRNA in HistoGene® samples. RT-PCR performed on RNA from 500 cells captured from mouse liver and kidney. Equal quantities of cDNA analyzed with three primer sets: elongation factor 1-a (EF-1a, high-abundance gene, ~3,000 copies/cell, 187 bp), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, medium-abundance gene, 300–3,000 copies/cell, 357 bp), and protein phosphatase 1 (PP1, low-abundance gene, <300 copies/cell, 498 bp). Samples were run on a 6% acrylamide gel and stained using SYBR® Gold Nucleic Acid Gel Stain (Life Technologies). M: molecular weight markers. 1: kidney EF-1a. 2: liver EF-1a. 3: kidney GAPDH. 4: liver GAPDH. 5: kidney PP1. 6: liver PP1. 7: liver EF-1a no RT control. 8: no RT template control.

Obtain High-Quality RNA from Many Tissue Types
The HistoGene® kit has been validated by examining RNA integrity on an expanding list of tissue types. All tissues tested to date using the HistoGene® kit have yielded high-quality RNA (Table 1).

Table 1. Tissues Validated with the HistoGene® Kit.

The Arcturus® HistoGene® LCM Immunofluorescence Staining Kit is designed to retrieve high-quality nucleic acids from immunofluorescently stained, frozen tissue using a quick 15-minute process. The kit provides reagents and slides for convenient and reliable immunofluorescent staining as well as protocols that are streamlined and optimized to maintain nucleic acid quality.

Each kit provides enough material for staining 32 slides and includes:

• HistoGene® Buffer A (4 mL)
• HistoGene® Buffer B (60 mL)
• HistoGene® Cy®3 Streptavidin (60 μL)
• 72 HistoGene® LCM Immunofluorescence Staining Slides
• User guide

Brilliant High-Contrast Label Intensity
The HistoGene® LCM Immunofluorescence Staining Kit employs a biotin–avidin system with Cy®3 dyes, resulting in exceptionally good staining intensity and specificity. The staining kit is used with a biotinylated monoclonal primary antibody against an antigen of choice provided by the user, and Cy®3 conjugated streptavidin is included in the kit (Figures 2).

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Figure 2. Identify target cells using the HistoGene® LCM Immunofluorescence Staining Kit. (A) Human foreskin tissue stained with anti-CD1a antibody. (B) Human jejunum tissue stained with anti-pan-cytokeratin antibody. (C) Mouse lacrimal gland inflammatory infiltrate stained with anti-CD4 antibody. (D) Mouse small intestine tissue stained with anti-MAdCAM-1 antibody.

Obtain High-Quality RNA from Many Tissue Types
The HistoGene® LCM Immunofluorescence Staining Kit has been validated by examining RNA integrity on an expanding list of tissue types. All tissues tested to date using this kit have yielded quality RNA (Table 2).


Table 2. Validated tissue-antibody sets.

The Arcturus® Paradise® PLUS FFPE LCM Staining Kit offers a simple solution developed to stain formalin-fixed, paraffin-embedded (FFPE) tissues before proceeding with laser capture microdissection (LCM) and downstream molecular analysis. FFPE samples introduce unique challenges for gene expression profiling, including chemical modification and fragmentation of RNA molecules. Archiving of FFPE samples adds to these challenges through increased RNA degradation over time.