Pure Human Normal Hepatocytes Isolated by LCM from FFPE Sample

Description

Sample Basic Information

Sample ID	Organ	Pathology Diagnosis	Gender	Age	Grade	TMN	IHC Data	Sample Format
08005A	Human Liver	Human liver cancer (HuPS-08005T) adjacent normal tissue	Male	34	N/A	N/A	Ki67 IHC	FFPE

Tissue H&E Staining and IHC images

H&E Stain

Ki67 IHC

Flowchart of LCM  for The Product Preparation

1. FFPE samples are cut at 6 microns and mounted onto pen membrane glass slides and let the slides dry for 2 hours at room temperature.
2. After deparaffinization and rehydration, quick H&E staining is performed.
3. Arcturus XT Laser Microdissection System is used for LCM. Microdissection is performed using IR and UV laser and CapSure Macro LCM Cap.
4. The microdissected cells are on the film of Macro LCM Cap. The total dissected cells may be varied between 10,000 to 50,000 cells which depend on  tissue type. 
5. Customers can be isolated DNA, RNA or protein from LCM Caps for the downstream applications.

Quality Control

• The tissue H&E staining slides are examined by certified pathologists. Pathological re-confirmation report is generated and digital image captured. 
• LCM is supervised by our certified pathologist.
• Three example LCM images including Before, After LCM, and LCM cap images from LCM tissue slides are taken to demonstrate the dissected cells.

Product Format and Size

• Dissected cells on the film of Macro LCM Caps
• 2 LCM Macro caps

Extraction Protocol for DNA, RNA and Protein from LCM Caps

• For DNA extraction, you can use Arcturus® PicoPure® DNA Extraction Kit (Thermo Fisher, KIT0103)
• For RNA isolation, you can use Arcturus® Paradise® PLUS FFPE RNA Isolation Kit (Thermo Fisher, Cat# KIT03121)
• For protein extraction, you can follow the protocol from Longuespée R, et al. A laser microdissection-based workflow for FFPE tissue microproteomics: Important considerations for small sample processing, Methods;2016, 104: 154-162

Applications

• The  pure cell population on the film of caps can be used for extraction of DNA, RNA/miRNA or protein for RT-PCR, cDNA array, sequencing, epegenectics, proteomics and antibody array, etc. 
• The product is only used for biomedical research. It is not used for clinical diagnosis.

Storage

        Store at -80ºC 

References

1. Longuespée R, et al. A laser microdissection-based workflow for FFPE tissue microproteomics: Important considerations for small sample processing, Methods;2016, 104: 154-162
2. Cai J, et al. The Use of Laser Microdissection in the Identification of Suitable Reference Genes for Normalization of Quantitative Real-Time PCR in Human FFPE Epithelial Ovarian Tissue Samples. PLoS ONE 2014, 9(4): e95974. 
3. Drummond ES, et al. Proteomic analysis of neurons microdissected from formalin-fixed, paraffin-embedded Alzheimer’s disease brain tissue.Sci Rep. 2015;5:15456
4. Bockmeyer CL, et al. Recommendations for mRNA analysis of micro-dissected glomerular tufts from paraffin-embedded human kidney biopsy samples. BMC Mol Biol. 2018;19(1):2
5. Murray, Graeme I. (Ed.). Laser Capture Microdissection (Methods and Protocols), © 2018

Product Data Sheet (PDF)

LCM-C08005A Data Sheet