Stellaris RNA FISH 프로토콜은 간단하며 아래 이미지와 같이 네 가지 주요 단계로 구성됩니다.
일부 샘플 유형의 경우 전체 프로세스를 하루 이내에 완료할 수 있습니다.
Step A: Prepare a sample
A sample is grown, adhered to, or sectioned onto a #1 cover glass and permeabilized with alcohol. No protease is required!
Note: Variations in sample preparation between different organisms and sample types are covered in our online Stellaris protocols.
Step B: Hybridize probes
Hybridization can be completed in 4 to 16 hours at +37 °C in a standard laboratory incubator.
Step C: Wash the sample
Removal of the Hybridization Buffer and the addition of wash buffers with short incubation periods can remove excess probes. The total time for this step is 1-1.5 hours.
Step D: Image sample
The sample is ready for imaging using a standard fluorescence microscope. We recommend using a wide-field fluorescence microscope though other imaging platforms can be used with Stellaris RNA FISH probes. Image processing is done using available online tools which convert Z-stack images to a maximum-intensity projection image.