At the collection site

• Mix an equal volume of 50% ethanol or isopropyl alcohol with the urine sample.

At the laboratory

• Upon reception of the sample at the laboratory, add the ImmunoCyt™ /uCyt+™ fixative solution.
• Urine samples are kept between 2ºC and 8 ºC for a maximum period of 7 days from the collect date.

Control slides preparation

• Apply positive and negative controls cell suspension (included in the kit) on slides and fix with spray.

Filtration of sample

• Filter urine using a syringe through a polycarbonate filter mounted on a filter holder.
• The monolayer liquid-based preparation methods have not been validated by Scimedx. It is the responsibility of the laboratory to validate any method other than that which is suggested in the product package insert.

Cell transfer

• Remove the filter and place it on a slide. This transfers the cells on the slide; fix with a spray.
   

Pretreatment of slides

• Immerse slides in eight baths (ethanol 80%, 70%, 50%, deionized water and hematoxylin + 4% acetic acid).

Immunoreaction

Incubate slides in humid chamber with:

• blocking solution for 15 minutes;
• antibody solution for 60 minutes.

Mounting

• Rinse slides in the phosphate buffer solutions.
• Put one drop of mounting medium on slide and place a cover slip on it.
• Keep the slides protected from light between 2ºC and 8ºC for maximum period of 7 days until microscope reading.

Positive control

Negative control

Color of background (20X)

Positive

Presence of one confirmed red or green positive cell.  If less than 5 positive cells are counted, it is recommended to evaluate a second sample.

Negative

Absence of fluorescence on a slide with >500 epithelial cells.  Must obtain an average of one cell per microscopic field with the 20X objective on brightfield

Inadequate

Absence of fluorescence on a slide with <500 epithelial cells