I have been using in-house fabricated cDNA microarrays to study differential gene expression of approximately 100 human genes using custom-made cDNA microarrays. The great advantage of glass microarrays over nylon membranes is the low sample volume required, typically 10-20 µl, which allows high probe concentrations. After labeling, samples are applied to the microarray slide, covered with a glass coverslip and incubated in a humidified hybridization chamber at 65°C. Because of the low hybridization volume, sample evaporation is a serious concern. If the hybridization chamber does not seal well, the sample will evaporate and the microarray will have to be discarded. I have run into this problem using other hybridization chambers. The gasket of the first hybridization chamber I tried became leaky after less than 20 hybridizations which led to sample evaporation and/or flooding of the chamber. On another occasion I had a cover break because it could not withstand the force of the screws used to fasten it. Searching for a better product, I came across GeneMachine’s Hybchamber from Genomic Solutions. I have been using it for more than a year now and have never had any evaporation problems. The gasket and screws are still intact after more than 100 hybridizations. In addition, the hybridization chamber is easy to assemble and disassemble with no other tools required.
Genemachines’ HybChamber is designed to hold up to two slides, a feature that I appreciate. Slides are placed array-side up into each slide compartment along with about 10µl 1x hybridization solution. The hybridization solution goes into a reservoir next to the slide compartment. Next the chamber is sealed with a transparent cover and the butterfly screws are fastened tight. To keep temperature, the chamber is then placed into a water bath or other temperature controlled oven according to the users protocol. After hybridization, the unit is easily disassembled by unscrewing the butterfly screws.
So far, my experience with these chambers has been positive. The sturdy construction has given us consistently good performance. The units are well designed and can be stacked in a hybridization oven, which ensures uniform hybridization conditions among multiple cassettes. Also, the butterfly screws used to hold the cover on the chamber make fastening and unscrewing easy. But all of this does not come cheap. These hybridization chambers are high quality but they are more expensive than some other chambers on the market. Also, the transparent cover that these chambers come with makes inspection easy, but because light might damage the fluorescent samples, the hybridization oven needs to be protected from light in some way (e.g. covering it in aluminum foil).
Overall, this is a very robust and durable hybridization chamber. It has been very well designed and is easy to use. It is my first choice over other hybridization chambers.
Martin Bilban, MSc
PhD Student
Department of Cell Biology
The Scripps Research Institute