Histosonda®

Listed here are some common reasons for unexpected results but if these do not satisfy your query we will be happy to answer any further questions at This e-mail address is being protected from spambots. You need JavaScript enabled to view it

A cloudy precipitate remains in the HistoSonda® tube after reconstituting:

Pipette up and down to break up the precipitate. Vortex the tube and centrifuge for 10 seconds. If unusually, the precipitate still remains, you can cover the tissue section with the entire solution including the precipitate, using a pipette tip to spread evenly. This will not adversely affect the hybridization process or results. After hybridization, wash vigorously in PBS to remove any trace of precipitate. Alternatively, the precipitate may be dissolved as stated in the protocol, by introducing the tu be in an incubator at 62°C for 15mins prior to use.

Intense background is observed:

Heat Treatment - If the slides have been heat treated it is possible that the duration was too long. Decrease the time in the microwave slightly and remember to remove the slides and transfer them to room temperature water immediately once uniform boiling is observed.

Incubations - Probe or other incubation solution may have dried to tissue section. Use a individual slide chambers or humid chamber for all incubations making sure that it is well c1osed. Check slides after each incubation to be sure that solutions have not dried, and wash tissue sections well.

Antibody Dilution - Primary antibody dilution may be too concentrated. Refer to manufacturers instructions for guidance or for optimal results, use the antibody supplied by CENBIMO which is prepared at an optimum working dilution.

No labelling observed:

Protocol may have not been followed correctly. Check for omission of steps.

Tissue is negative for this probe.

Weak labelling observed:

Primary antibody dilution may be too weak. Refer to manufacturers instructions for guidance or for optimal results, use the antibody supplied by CENBIMO which is prepared at an optimum working dilution.

Tissue is expressing target RNA in small quantities:

Insufficient incubation with Proteinase K. Some tissues types may need a longer deproteinization period.

Eosinophils have been labeled positive:

Heat treatment of tissue sections may either have been omitted or insufficient. Increase the time in the microwave or boiling water slightly and remember not to remove slides until you see uniform boiling (bubbles rising from the bottom of the wash bath), however also take into account that increasing the time more than necessary will result in background labelling.

Unexpected results when using Bone Marrow sections:

The variable nature of bone marrow processing can affect the deproteinization efficiency of Proteinase K. If you observe weak or no labelling when using bone marrow sections we advise to increase the time of deproteinization until suitable results are obtained.