Mc63 camera system

Stool samples of children, which tested positive or probable for Giardia and/or Cryptosporidium by conventional microscopy and/or ICT, were further assayed by direct fluorescent antibody test (DFAT) for confirmation. Briefly, 5 μL of concentrated (as described above) stool samples were placed on welled slides. Smears were air-dried, methanol fixed, and stained with fluorescein-labeled mouse monoclonal antibodies (Crypto/Giardia Cel; Cellabs, Sydney, Australia). Samples were examined on a Zeiss fluorescence microscopy (Carl Zeiss, Oberkochen, Germany) equipped with a MC63 camera system (Carl Zeiss, Oberkochen, Germany) at ×400 magnification. Known positive and negative controls from clinical specimens submitted to our laboratory for diagnosis were routinely included in each sample batch.

A direct fluorescent antibody test was used to confirm presumptive microscopy/ICT positive results. Briefly, stool samples (~1 g) were processed using the concentration system PARASEP Midi^® (Grifols Movaco, Barcelona, Spain) according to the manufacturer’s instructions. Five μL of concentrated faecal material were placed on welled slides. Smears were air-dried, methanol fixed, and stained with fluorescein-labelled mouse monoclonal antibodies directed against Cryptosporidium oocysts and Giardia cysts (Crypto/Giardia Cel, Cellabs, Sydney, Australia). Samples were examined on a Zeiss fluorescence microscopy equipped with a MC63 camera system at 400x magnification. Known positive and negative controls were routinely included in each sample batch.