Tcs sp5 inverted laser scanning microscope

Manufactured by Leica

Sourced in Germany

The TCS SP5 is an inverted laser scanning microscope manufactured by Leica. It is designed for advanced fluorescence imaging and analysis. The system utilizes multiple laser lines and sensitive detectors to capture high-resolution images of biological samples.

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Lab products found in correlation

Chambered cell culture cover glasses, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

TCS SP5 inverted laser scanning confocal microscope TCS SP5 inverted microscope TCS SP5 scanning microscope TCS SP5 microscope Inverted SP5 microscope Laser scanning microscope TCS SP5 SP5 Inverted SP5 microscope Inverted microscope

4 protocols using tcs sp5 inverted laser scanning microscope

Mitochondrial and Nuclear Imaging Assay

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Cells were seeded
at 100,000 cells/well in a four-well chambered cell culture cover
glass and left to grow for 24 h. They were then transfected with CellLight
Mitochondria-GFP BacMam 2.0 and left to incubate overnight. The cells
were then incubated with fTPP@(DCA₅-UiO-66) for 2 h, after
which the media was removed, and the wells washed twice with PBS.
Five μM of DRAQ5 solution in media was added to the cells and
left to incubate at room temperature in the dark for 5–30 min.
The cells were then analyzed directly without further washing using
a TCS SP5 inverted laser scanning microscope (Leica, Germany). An
argon laser was used to visualize GFP-stained mitochondria (excitation
at 488 nm and emission filter set at 505–555 nm). To visualize
the stained nucleus, a helium–neon laser was used (excitation
at 633 nm and emission filter set at 650–700 nm). fTPP@(DCA₅-UiO-66) was excited using a diode laser emitting at 405 nm.
Images were taken sequentially. Fluorescent images of cells were acquired
as described above. Images were then merged and converted to 8-bit
RGB format using ImageJ software.

Haddad S., Abánades Lázaro I., Fantham M., Mishra A., Silvestre-Albero J., Osterrieth J.W., Kaminski Schierle G.S., Kaminski C.F., Forgan R.S, & Fairen-Jimenez D. (2020). Design of a Functionalized Metal–Organic Framework System for Enhanced Targeted Delivery to Mitochondria. Journal of the American Chemical Society, 142(14), 6661-6674.

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Multispectral Imaging of Cellular Uptake

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Cells were seeded on 1.8 cm 2 chambered cell culture cover glasses (Nunc, UK) at a density of 3 x 10 4 cells/well. Cells were grown for 24-48 hours and then incubated for 2 hours with 100 µg/ml transferrin-AF488, 2 µM lacCer-BODIPY or 1 mg/ml fluorescein isothiocyanatedextran (150 kDa) in the presence of 250 µg/ml PP-50/AF647. Subsequently, cells were incubated for 15 minutes with 5 µg/ml Hoechst 33342; this permeable dye is used to evaluate nuclear morphology as it emits fluorescence when bound to DNA. Cells were washed twice with PBS and extracellular labelled molecules were quenched using 0.4 % trypan blue [20] (link). Cells were washed again with PBS and left in growth media. Cells were analysed using a TCS SP5 inverted laser scanning microscope (Leica, Germany). An argon laser was used to visualise transferrin-AF488, lacCer-BODIPY, and fluorescein isothiocyanate-dextran (emission at 488 nm and an emission filter set at 505-555 nm). To visualise PP-50/AF647, a helium neon laser was used (emission at 633 nm and emission filter set at 650-700 nm). H33342 stained nuclei were excited using a diode laser emitting at 405 nm. Images were taken sequentially.

Mercado S.A., Orellana-Tavra C., Chen A, & Slater N.K. (2016). The intracellular fate of an amphipathic pH-responsive polymer: Key characteristics towards drug delivery. Materials science & engineering. C, Materials for biological applications, 69.

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Multicolor Organelle Tracking in Cells

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Cells were seeded on 1.8 cm 2 chambered cell culture cover glasses (Nunc, UK) at a density of 3 x 10 4 cells/well and grown for 24-48 hours. Cells were then transduced with CellLight reagents according to the manufacturer's instructions. Briefly, cells were incubated for 16 hours with 30 µl of CellLight solution with baculovirus containing either early endosomes Rab5a-GFP, late endosomes Rab7a-GFP or lysosomes Lamp-1-GFP. After this period, cells were washed twice with PBS, incubated with 250 µg/ml PP-50/AF647 for 2 and 24 hours and finally incubated for 15 minutes with 5 µg/ml H33342. Cells were washed twice with PBS and extracellular labelled molecules were quenched using 0.4 % trypan blue. Cells were washed again with PBS and left in growth media. Cells were analysed using a TCS SP5 inverted laser scanning microscope (Leica, Germany). An argon laser was used to visualise the GFP labelled early endosomes, late endosomes and lysosomes (excitation at 488 nm and emission filter set at 505-555 nm). To visualise PP-50/AF647, a helium neon laser was

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Trehalose Uptake and Cell Viability Imaging

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Cells were seeded on 1.8 cm 2 chambered cell culture cover glasses (Nunc, UK) at a density of 3 x 10 4 cells/well and grown for 24-48 hours. Cells were then incubated with 50 mM FITCtrehalose in the presence or absence of 250 µg/ml PP-50 for 2, 4, and 6 hours. FITCtrehalose and PP-50 solutions were prepared in DMEM. Cells were washed twice with PBS and extracellular labelled molecules were quenched using 0.4 % trypan blue [30] (link). Cells were washed again with PBS and left in growth media. Cells were incubated for 15 minutes with 5 µg/ml PI and 5 µg/ml H33342 then incubated in growth media and analysed using a TCS SP5 inverted laser scanning microscope (Leica, Germany). An argon laser was used to visualise FITC-trehalose (emission at 488 nm and an emission filter set at 505-555 nm). To visualise propidium iodide, a helium neon laser was used (emission at 633 nm and emission filter set at 650-700 nm). H33342 stained nuclei were excited using a diode laser emitting at 405 nm. Images were taken sequentially.

Mercado S.A, & Slater N.K. (2016). Increased cryosurvival of osteosarcoma cells using an amphipathic pH-responsive polymer for trehalose uptake. Cryobiology, 73(2).

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