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Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

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An improved fixation technique for fluorescence in situ hybridization for preimplantation genetic diagnosis

D I Dozortsev¹, K T McGinnis

¹Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Hutzel Hospital, Wayne State University, Detroit, MI 48201, USA. aa4421@wayne.edu

Fertility and Sterility

|July 27, 2001

Abstract

Objective:

To improve existing preimplantation genetic diagnosis fixation techniques.

Design:

Prospective randomized in vitro study.

Setting:

Academic medical center.

Patient(S):

None.

Intervention(S):

None.

Main Outcome Measure(S):

The intensity and clarity of fluorescence in situ hybridization (FISH) signals and the percentage of successfully fixed blastomeres.

Result(S):

The described fixation technique resulted in 100% fixation and 100% adequate FISH signals. Two conventional techniques resulted in 94% and 87% fixation, and in adequate FISH signals in 81% and 87%, respectively.

Conclusion(S):

This newly developed fixation technique simplifies the process of fixation of blastomeres for preimplantation diagnosis while essentially eliminating the possibility of losing a cell during fixation. It will hopefully allow more IVF programs to offer their patients preimplantation genetic diagnosis using the FISH technique.

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FISH - Fluorescent In-situ Hybridization

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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

In-situ Hybridization

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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