Mrm ccd camera, by Zeiss - Product details

1

Lysosomal and Calcium Imaging Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lysosomes were visualised in live cells in chamberslides (Ibidi) using
300 nM LysoTracker red or green (Life Technologies) in Dulbecco’s
modified phosphate buffered saline (DPBS) for 15-minutes at room temperature,
washed tree times with DPBS, and imaged using a Zeiss Axio Observer inverted
microscope with Colibri LED light source and Zeiss Mrm CCD camera with
Axiovision 4.8 software. Lysosomal area per cell was measured from LysoTracker
fluorescence images in ImageJ 1.50i and 1.52n11 (link)
using the analyse particles function. LysoTracker fluorescence was measured in
cells grown in Corning CellBIND 96-well plates (0.8x10^5 cells/well) using a
SpectraMax® Gemini microplate reader (Molecular Devices).
Ca²⁺ measurements were done as described12 (link) but with minor modifications for neurons, which
were loaded with 1 µM Fura-2, AM (Stratech) without Pluronic F-127. Cells
were imaged in Hank’s balanced salt solution (HBSS; 1 mM HEPES pH7.4, 10
µM CaCl₂ and 1 mM MgCl₂) using a Zeiss Axiovert 35
microscope with Cairn Optospin filter exchanger, Orca Flash 4.0 sCMOS camera and
MetaFluor 7.10 software. For all experiments, ionomycin (Merck, 2 μM) was
added to clamp intracellular Ca²⁺ stores followed by 500 µM
Gly-Phe-β-naphthylamide (GPN, Abcam) to release lysosomal Ca^{2+12 (link)}.

Cook S.R., Badell-Grau R.A., Kirkham E.D., Jones K.M., Kelly B.P., Winston J., Waller-Evans H., Allen N.D, & Lloyd-Evans E. (2020). Detrimental effect of zwitterionic buffers on lysosomal homeostasis in cell lines and iPSC-derived neurons. AMRC open research, 2, 21.

+ Open protocol

+ Expand

2

Inhibition of Candida Biofilm Formation by Lactobacillus

Check if the same lab product or an alternative is used in the 5 most similar protocols

To evaluate the effects of L. casei Shirota on the yeast-to-hyphae transition and initial biofilm formation of Candida spp., cocultures of L. casei/Candida were grown in 96-well flat-bottom polystyrene plates. The experiment was carried out using clinical isolates and the reference C. albicans strains ATCC 90028 and SC5314. In brief, 100 μL/well RPMI 1640 medium was supplied with serum 10% (v/v) and inoculated with a cell suspension of 1 × 10⁶ yeast and 1 × 10⁸ bacteria per milliliter. The plates were incubated at 37 °C for 24 h to obtain early biofilms. Images were captured with either a Nikon Eclipse E200 or an Mrm CCD camera (Carl Zeiss GmbH), and the microscope and camera were jointly operated for image capture by Zen 2012 software with 10× and 40× objectives.

Paniágua A.L., Correia A.F., Pereira L.C., de Alencar B.M., Silva F.B., Almeida R.M, & de Medeiros Nóbrega Y.K. (2021). Inhibitory effects of Lactobacillus casei Shirota against both Candida auris and Candida spp. isolates that cause vulvovaginal candidiasis and are resistant to antifungals. BMC Complementary Medicine and Therapies, 21, 237.

+ Open protocol

+ Expand

3

Lysosomal and Calcium Imaging Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lysosomes were visualised in live cells in chamberslides (Ibidi) using
300 nM LysoTracker red or green (Life Technologies) in Dulbecco’s
modified phosphate buffered saline (DPBS) for 15-minutes at room temperature,
washed tree times with DPBS, and imaged using a Zeiss Axio Observer inverted
microscope with Colibri LED light source and Zeiss Mrm CCD camera with
Axiovision 4.8 software. Lysosomal area per cell was measured from LysoTracker
fluorescence images in ImageJ 1.50i and 1.52n11 (link)
using the analyse particles function. LysoTracker fluorescence was measured in
cells grown in Corning CellBIND 96-well plates (0.8x10^5 cells/well) using a
SpectraMax® Gemini microplate reader (Molecular Devices).
Ca²⁺ measurements were done as described12 (link) but with minor modifications for neurons, which
were loaded with 1 µM Fura-2, AM (Stratech) without Pluronic F-127. Cells
were imaged in Hank’s balanced salt solution (HBSS; 1 mM HEPES pH7.4, 10
µM CaCl₂ and 1 mM MgCl₂) using a Zeiss Axiovert 35
microscope with Cairn Optospin filter exchanger, Orca Flash 4.0 sCMOS camera and
MetaFluor 7.10 software. For all experiments, ionomycin (Merck, 2 μM) was
added to clamp intracellular Ca²⁺ stores followed by 500 µM
Gly-Phe-β-naphthylamide (GPN, Abcam) to release lysosomal Ca^{2+12 (link)}.

Cook S.R., Badell-Grau R.A., Kirkham E.D., Jones K.M., Kelly B.P., Winston J., Waller-Evans H., Allen N.D, & Lloyd-Evans E. (2020). Detrimental effect of zwitterionic buffers on lysosomal homeostasis in cell lines and iPSC-derived neurons. AMRC open research, 2, 21.

+ Open protocol

+ Expand

4

Imaging Fluorescent Samples in Optimal Conditions

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ultrathin sections were imaged for fluorescent signal in two ways, depending on the quality of resin infiltration and embedding. Where embedding was less than optimal, it proved necessary to image sections hydrated to eliminate reflections from holes and tears in the sections. For this, grids were mounted between a glass slide and coverslip using phosphate buffered glycerol, pH 8.6. However, in optimal conditions this step was unnecessary, and the fluorescent signal could be directly collected from grids placed upon glass slides. Images were collected with the standard widefield epifluorescence microscope using 40× or 100× air objectives (EC Plan-Neofluar 40×/0.75; EC Epiplan-Neofluar 100×/0.75) and an MRm CCD camera (Zeiss, Cambridge).

Peddie C.J., Blight K., Wilson E., Melia C., Marrison J., Carzaniga R., Domart M.C., O׳Toole P., Larijani B, & Collinson L.M. (2014). Correlative and integrated light and electron microscopy of in-resin GFP fluorescence, used to localise diacylglycerol in mammalian cells. Ultramicroscopy, 143(100), 3-14.

+ Open protocol

+ Expand

5

Capsule Induction and Cell Division Analysis

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Macroscopic and microscopic cell morphology was observed in two capsule-inducing media: CO₂-independent medium [CIM; (Ost et al., 2015 (link))] and 10-fold diluted Sabouraud in 50 mM MOPS [Sab/MOPS; (Zaragoza and Casadevall, 2004 (link))]. CIM was also used to document cell division by bright field, time-lapse microscopy. To do that, cells were seeded onto a well of a 96-well plate, at a density of a thousand cells per well in 100 μL of CIM. Pictures were taken every 30 min in the same microscope as above, over a course of 48 h at 37°C using a 10X objective. A sample of each culture was also applied to a microscope slide for inspection at 63X (Plan-Apochromatic 63X/1.40 DIC; Carl Zeiss GmbH) using differential interference contrast (DIC). Images were captured with an Mrm CCD camera (Carl Zeiss GmbH) and the whole microscope and camera were operated for image capture by the Zen 2012 software.
Cells from these cultures were also sonicated using a QSonica Q700 Sonicator equipped with a 1.6 mm tip. The settings were 10% amplitude for intervals of up to 1 min, when all cells were destroyed. At intermediate times, aliquots were inspected under bright field microscopy to assess whether cell clumps had dispersed.

Paes H.C., Derengowski L.D., Peconick L.D., Albuquerque P., Pappas GJ J.r., Nicola A.M., Silva F.B., Vallim M.A., Alspaugh J.A., Felipe M.S, & Fernandes L. (2018). A Wor1-Like Transcription Factor Is Essential for Virulence of Cryptococcus neoformans. Frontiers in Cellular and Infection Microbiology, 8, 369.

+ Open protocol

+ Expand

6

Live Cell Imaging of HeLa Cells Treated with RHL

Check if the same lab product or an alternative is used in the 5 most similar protocols

HeLa cells treated with RHL 160 µmol/l were used for live cell imaging for 24 h by placing the culture dishes onto a sample stage within a heated chamber (37°C). Live imaging was performed using an ImageXpress Micro XLS Widefield High-Content Analysis system (Molecular Devices, LLC, Sunnyvale, CA, USA). Images were captured at 40×` with 10 msec exposure times in 5 min intervals using an MRm CCD camera (Carl Zeiss AG, Oberkochen, Germany), and different Z sections were projected using the softWoRx^® Suite (version 2.0; Applied Precision, Inc., Mississauga, ON, Canada).

Liu J., Zhen Y.Z., Cui J., Hu G., Wei J., Xu R., Tu P, & Lin Y.J. (2018). Dynamic influence of Rhein lysinate on HeLa cells. International Journal of Oncology, 53(5), 2047-2055.

+ Open protocol

+ Expand

Mrm ccd camera

Lab products found in correlation

Close spelling variants of this product

6 protocols using mrm ccd camera

Lysosomal and Calcium Imaging Protocol

Inhibition of Candida Biofilm Formation by Lactobacillus

Lysosomal and Calcium Imaging Protocol

Imaging Fluorescent Samples in Optimal Conditions

Capsule Induction and Cell Division Analysis

Live Cell Imaging of HeLa Cells Treated with RHL

About PubCompare

Ready to get started?