Micromanipulation of single cells from tissue imprints is an alternative to laser-assisted microdissection.
The characterization and analysis of single cells by molecular biological methods such as the polymerase chain reaction (PCR) is of increasing interest in biomedical research. Different techniques have been developed to obtain single cells from solid tissue. Currently, the most frequently used technique is laser-assisted microdissection (LAM). However, LAM of tissues cannot exclude contamination of the targeted cells by underlying cell fragments. Moreover, this technique can only be performed if a laser microscope is available. Thus, we developed a method to obtain single cells of fresh solid tissue by the simple technique of tissue imprints. After immunostaining of the imprints, single cells were transferred to a reaction tube using a 27-gauge needle guided by a mechanical micromanipulator. Consequently, we used these cells in a single cell PCR.
Brauns
Tilmann C
TC
Department of Dermatology, Venerology and Allergology, University of Essen, Essen, Germany. tilmannbrauns@aol.com
Goos
Manfred
M
D001483:Base Sequence; D003581:Cytodiagnosis / Q000295:instrumentation / Q000379:methods*; D005260:Female; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D008846:Micromanipulation / Q000379:methods*; D008969:Molecular Sequence Data; D009182:Mycosis Fungoides / Q000235:genetics / Q000473:pathology; D016133:Polymerase Chain Reaction; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics; D017422:Sequence Analysis, DNA; D012867:Skin / Q000473:pathology*; D012878:Skin Neoplasms / Q000235:genetics / Q000473:pathology
D016692:Receptors, Antigen, T-Cell, gamma-delta