Tcs sp8 aobs microscope

Light microscopy observations of live cells of Gambierdiscus were carried out under at ×1000 magnification with a Zeiss Imager.A2 microscope equipped Nomarski differential interference contrast (D.I.C.) optics. Cells fixed with glutaraldehyde (5% final concentration) were stained with DAPI (4',6' diamino-2-phenylindole) and observed at ×1000 magnification under an Olympus BX51 epifluorescence microscope equipped with an Olympus DP72 camera (Olympus, Tokyo, Japan). For plate pattern identification, the cells from cultures were dissected, squashed by gently pressing the cover slip over them, and occasionally with the aid of sodium hypochlorite (2%) and observed at ×1000 magnification under an Olympus BX51 microscope. Cells fixed with glutaraldehyde were stained with both SYBR Green I (green emission) and Fluorescent Brightener 28 (blue emission) and observed at ×630 magnification with an inverted Confocal Leica TCS SP8 AOBS microscope. The nomenclature for the plate tabulation followed Litaker et al. [11 ]. Cell size was measured by light microscopy on 15–20 randomly selected living cells. Cell size was described as depth (ventral to dorsal distance), width (transdiameter), and length (apical to antapical axis measured in either ventral or dorsal view).

After allowing the animals to eliminate the excess of dye, they were transferred on glass slides with 4% agar pads and immobilized alive for microscopy analysis with 0.01% tetramisole hydrochloride (cat. n. T1512 Sigma-Aldrich^®). Confocal and epi-fluorescence (FITC filter) images were collected with Leica TCS SP8 AOBS microscope using a 40x objective. For persistence analysis, the animals were collected and observed after 24, 48 and 72 h from the treatment.