axiovert 200m wide-field fluorescence microscope  (Carl Zeiss)

Journal: Pharmaceutics

Article Title: Development of Breast Cancer Spheroids to Evaluate Cytotoxic Response to an Anticancer Peptide

doi: 10.3390/pharmaceutics13111863

Figure Lengend Snippet: Evaluation of the MDA-MB-231 spheroid formation. ( A ) Spheroid diameter (µm) daily monitoring over 14 days measured with a widefield fluorescence microscope. ( B ) Spheroid cell metabolic activity daily evaluation over 14 days by measuring the fluorescence emission of resorufin with a microplate reader. ( C ) Representative images of parameters evaluated on days 7 and 14 at 2000 cells/well with a confocal point-scanning Zeiss LSM 880 microscope. In the apoptosis determination, apoptotic cells are marked in green; in the oxidative stress measurement, reactive oxygen species (ROS) are marked in yellow; and in the cell viability/mortality evaluation, intact cells (live) are marked in green and permeabilized cells (dead) in red. In all experiments, cell nucleus is marked in blue. Scale bar = 200 µm. Graphs represent at least three biological repeats.

Article Snippet: During spheroid optimization, diameter and morphology were monitored daily from day 1 (i.e., the day after addition of cellular matrix) to day 14, using the wide field fluorescence microscope Zeiss Axiovert 200M (Carl Zeiss, Oberkochen, Germany) equipped with an EC Plan-Neofluar ×10 dry objective (0.30 numerical aperture) and a Leica DFC450 camera.

Techniques: Fluorescence, Microscopy, Activity Assay

Journal: Pharmaceutics

Article Title: Development of Breast Cancer Spheroids to Evaluate Cytotoxic Response to an Anticancer Peptide

doi: 10.3390/pharmaceutics13111863

Figure Lengend Snippet: Evaluation of the BT-20 spheroid formation. ( A ) Spheroid diameter (µm) daily monitoring over 14 days measured with a widefield fluorescence microscope. ( B ) Spheroid cell metabolic activity daily evaluation over 14 days by measuring the fluorescence emission of resorufin with a microplate reader. ( C ) Representative images of parameters evaluated on days 7 and 14 at 5000 cells/well with a confocal point-scanning Zeiss LSM 880 microscope. In the apoptosis determination, apoptotic cells are marked in green; in the oxidative stress measurement, reactive oxygen species (ROS) are marked in yellow; and in the cell viability/mortality evaluation, intact cells (live) are marked in green and permeabilized cells (dead) in red. In all experiments, cell nucleus is marked in blue. Scale bar = 200 µm. Graphs represent at least three biological repeats.

Article Snippet: During spheroid optimization, diameter and morphology were monitored daily from day 1 (i.e., the day after addition of cellular matrix) to day 14, using the wide field fluorescence microscope Zeiss Axiovert 200M (Carl Zeiss, Oberkochen, Germany) equipped with an EC Plan-Neofluar ×10 dry objective (0.30 numerical aperture) and a Leica DFC450 camera.

Techniques: Fluorescence, Microscopy, Activity Assay

Journal: Pharmaceutics

Article Title: Development of Breast Cancer Spheroids to Evaluate Cytotoxic Response to an Anticancer Peptide

doi: 10.3390/pharmaceutics13111863

Figure Lengend Snippet: Evaluation of the BT-474 spheroid formation. ( A ) Spheroid diameter (µm) daily monitoring over 14 days measured with a widefield fluorescence microscope. ( B ) Spheroid cell metabolic activity daily evaluation over 14 days by measuring the fluorescence emission of resorufin with a microplate reader. ( C ) Representative images of parameters evaluated on days 7 and 14 at 5000 cells/well with a confocal point-scanning Zeiss LSM 880 microscope. In the apoptosis determination, apoptotic cells are marked in green; in the oxidative stress measurement, reactive oxygen species (ROS) are marked in yellow; and in the cell viability/mortality evaluation, intact cells (live) are marked in green and permeabilized cells (dead) in red. In all experiments, cell nucleus is marked in blue. Scale bar = 200 µm. Graphs represent at least three biological repeats.

Article Snippet: During spheroid optimization, diameter and morphology were monitored daily from day 1 (i.e., the day after addition of cellular matrix) to day 14, using the wide field fluorescence microscope Zeiss Axiovert 200M (Carl Zeiss, Oberkochen, Germany) equipped with an EC Plan-Neofluar ×10 dry objective (0.30 numerical aperture) and a Leica DFC450 camera.

Techniques: Fluorescence, Microscopy, Activity Assay

Journal: Pharmaceutics

Article Title: Development of Breast Cancer Spheroids to Evaluate Cytotoxic Response to an Anticancer Peptide

doi: 10.3390/pharmaceutics13111863

Figure Lengend Snippet: Confocal microscopy characterization of apoptosis, reactive oxygen species (ROS) production, and cell viability/mortality in spheroids after treatment with peptides. Apoptosis, ROS production, and cell viability/mortality of ( A – C ) MDA-MB-231, ( D – F ) BT-20, and ( G – I ) BT-474 spheroids, formed under optimized conditions, after treatment for 5 days with vCPP2319 and PepH3 were assessed using a confocal point-scanning Zeiss LSM 880 microscope. The analysis was performed by calculating the corrected total cell fluorescence (CTCF) of different images and CTCF normalized to control. Graphs represent at least triplicate biological repeats, and results are displayed as mean ± SD, where * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: During spheroid optimization, diameter and morphology were monitored daily from day 1 (i.e., the day after addition of cellular matrix) to day 14, using the wide field fluorescence microscope Zeiss Axiovert 200M (Carl Zeiss, Oberkochen, Germany) equipped with an EC Plan-Neofluar ×10 dry objective (0.30 numerical aperture) and a Leica DFC450 camera.

Techniques: Confocal Microscopy, Microscopy, Fluorescence, Control