Laser microdissection (LM) is a powerful tool for various molecular analyses providing pure samples for genomic, transcriptomic, and proteomic studies. Cell subgroups, individual cells, or even chromosomes can be separated via laser beam from complex tissues, visualized under the microscope, and used for subsequent molecular analyses. This technique provides information on nucleic acids and proteins, keeping their spatiotemporal information intact. In short, the slide with tissue is placed under the microscope, imaged by a camera onto a computer screen, where the operator selects cells/chromosomes based on morphology or staining and commands the laser beam to cut the specimen following the selected path. Samples are then collected in a tube and subjected to downstream molecular analysis, such as RT-PCR, next-generation sequencing, or immunoassay.
- Klíčová slova
- Cells, Chromosomes, Cytology, Histology, Microdissection,
- MeSH
- analýza jednotlivých buněk MeSH
- chromozomy MeSH
- genom * MeSH
- laserová záchytná mikrodisekce metody MeSH
- proteomika * MeSH
- Publikační typ
- časopisecké články MeSH
Laser capture microdissection is a relatively young method used both in biomedical sciences as in other studies of animal and vegetable tissues and cells. Current human medicine and its methods of investigation are based on both current established processes, and simultaneously there are new experimental approaches from molecular biology tested. In this context it is highly desirable that the studied tissue is homogenous and representative population of cells. For this purposes at the late 80s the method of laser capture microdissection (LCM) has been developed, the first publication dealing with this method was released even in 1996. In current databases of literature we are able to find hundreds of papers focused on LCM such a method or as a part of methodic approach of experiments whose results led to the improved knowledge of genetic and proteomic nature of various diseases. This knowledge is of great promise for successful targeted therapy in the future.
In this study, we investigated repetitive sequences localized on Y chromosomes. Repetitive DNA sequences represent a substantial part of the eukaryotic genome and, among them, a large portion comprises sequences repeated in tandem. Efficient and rapid isolation of repeat units is possible due to a laser microdissection technique used for Y chromosome separation, followed by polymerase chain reaction (PCR), cloning, and sequence analysis. We applied the derived repeat units to members of nine tribes within the Bovidae. Apart from the Y chromosomes of Bos taurus and Bubalus bubalis, where we used known sequences of repetition, the derived sequences were used as probes for fluorescent in situ cross-hybridization to members of the nine tribes of the Bovidae. We investigated the distribution of repeat units within the tribes and their localization on the Y chromosome. Sharing of sequence variants would indicate common descent, while the rapid horizontal evolution should allow discrimination between closely related species or subspecies.
- MeSH
- chromozom Y chemie genetika MeSH
- druhová specificita MeSH
- fluorescenční barviva chemie MeSH
- fylogeneze MeSH
- hybridizace in situ fluorescenční veterinární MeSH
- laserová záchytná mikrodisekce metody veterinární MeSH
- metafáze MeSH
- molekulární evoluce MeSH
- populační genetika metody MeSH
- repetitivní sekvence nukleových kyselin * MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA metody veterinární MeSH
- sekvenční seřazení MeSH
- sexuální faktory MeSH
- skot klasifikace genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot klasifikace genetika MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fluorescenční barviva MeSH
