Tcs sp8 high speed confocal microscope

Manufactured by Leica

The Leica TCS SP8 is a high-speed confocal microscope designed for advanced imaging applications. It offers a range of features, including a fast scanning system, high-sensitivity detectors, and a modular design that allows for customization to meet specific research needs. The core function of the TCS SP8 is to provide high-resolution, three-dimensional imaging of biological samples with minimal phototoxicity and photobleaching.

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Lab products found in correlation

Application suite x software, by Leica (5 mentions) Mitotracker green fm, by Thermo Fisher Scientific (2 mentions) Lysotracker green dnd 26, by Thermo Fisher Scientific (2 mentions) Er tracker green, by Thermo Fisher Scientific (2 mentions) Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions) Lysotracker deep red, by Thermo Fisher Scientific (1 mentions) Er tracker red, by Thermo Fisher Scientific (1 mentions) P35g 0 14 c, by MatTek (1 mentions)

Spelling variants of this product

TCS SP8 (3513 citations) SP8 confocal microscope (2556 citations) Confocal microscope (1146 citations) TCS SP8 microscope (350 citations) SP8 microscope (288 citations) Confocal SP8 (11 citations) Confocal TCS SP8 (5 citations) TCS SP8 Confocal/Multi-Photon high-speed upright microscope (2 citations)

7 protocols using tcs sp8 high speed confocal microscope

Intracellular Localization of Compound 1 in A549 Cells

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Approximately 2 ×
10⁵ A549 cells in cell culture medium (2 mL) were seeded
on a confocal dish and incubated overnight at 37 °C in a humidified
5% CO₂ atmosphere. After being rinsed with PBS, the cells
were incubated with 1 (4 μM) at 37 °C for
1 h. After that, the cells were stained with LysoTracker Deep Red
(Thermo Fisher Scientific, Inc., L12492) (0.1 μM for 30 min),
MitoTracker Red CMXRos (Thermo Fisher Scientific, Inc., M7512) (0.1
μM for 20 min), or ER-Tracker Red (Thermo Fisher Scientific,
Inc., E34250) (1 μM for 20 min) in a serum-free medium at 37
°C. The solutions were then removed, and the cells were rinsed
with PBS twice before being examined with a Leica TCS SP8 high-speed
confocal microscope equipped with a 488 nm laser, a 552 nm laser,
and a 638 nm laser. LysoTracker Deep Red was excited at 638nm, and
the fluorescence was monitored at 650–680 nm. MitoTracker Red
CMXRos and ER-Tracker Red were excited at 552 nm, and their fluorescence
was monitored at 590–620 nm. Compound 1 was excited
at 488 nm and its fluorescence was monitored at 500–600 nm.
The images were digitized and analyzed using a Leica Application Suite
X software.

Durán-Sampedro G., Xue E.Y., Moreno-Simoni M., Paramio C., Torres T., Ng D.K, & de la Torre G. (2023). Glycosylated BODIPY- Incorporated Pt(II) Metallacycles for Targeted and Synergistic Chemo-Photodynamic Therapy. Journal of Medicinal Chemistry, 66(5), 3448-3459.

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Imaging of Organelle Dynamics in HT29 Cells

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HT29 cells on glass-bottom dishes of 35 mm diameter (MatTek Corporation, P35G-0-14-C) were incubated in the culture medium with 4 (2 μM) for 10 min at 37 °C. After being washed twice with PBS, the cells were refed with 2 mL of the culture medium and incubated for 6 h, and then stained with 0.2 μM ER-Tracker green (Thermo Fisher Scientific Inc., E34251), 0.2 μM MitoTracker Green FM (Thermo Fisher Scientific Inc., M7514) or 2 μM LysoTracker Green DND-26 (Thermo Fisher Scientific Inc., L7526) in Hank's Balanced Salt Solution (HBSS) at 37 °C for 15, 15 and 30 min respectively. After being washed twice with PBS, the cells were refed with HBSS and observed using a Leica TCS SP8 high speed confocal microscope equipped with a 488 nm argon laser and a 633 nm laser. All the trackers were excited at 488 nm, and their fluorescence was monitored at 500–570 nm, while conjugate 4 was excited at 638 nm and its fluorescence was monitored at 650–760 nm.

Xue E.Y., Wong R.C., Wong C.T., Fong W.P, & Ng D.K. (2019). Synthesis and biological evaluation of an epidermal growth factor receptor-targeted peptide-conjugated phthalocyanine-based photosensitiser. RSC Advances, 9(36), 20652-20662.

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Visualizing Cytochrome C Uptake in HT29 Cells

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HT29 cells were seeded at a density of 1 × 10⁵ cells per 35-mm confocal dish and incubated for 24 h. The culture medium was then replaced with 20 µM cytochrome C conjugates, and the cells were incubated for an additional 4 h. Following treatment, the cells were washed with phosphate-buffered saline (PBS) three times and stained with 1 µg mL⁻¹ Hoechst 33,342 nuclear dye (Phygene) at 37 °C for 10 min. Cells were washed with PBS three times to remove excess dye. Cellular uptake of the cytochrome C conjugates was visualised using a Leica TCS SP8 high-speed confocal microscope equipped with solid-state lasers. The fluorescein was excited at 488 nm, and its fluorescence was monitored at 500–530 nm. The Hoechst 33,342 was excited at 405 nm, and its fluorescence was monitored at 415–480 nm. The images were digitised and analysed using Leica Application Suite X software.

Tang B., Lau K.M., Zhu Y., Shao C., Wong W.T., Chow L.M, & Wong C.T. (2024). Chemical Modification of Cytochrome C for Acid-Responsive Intracellular Apoptotic Protein Delivery for Cancer Eradication. Pharmaceutics, 16(1), 71.

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Cellular Uptake of Photosensitizer-Biomolecule Conjugates

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Approximately 2 × 10⁵ HT29, A549, and HepG2 cells in the corresponding culture medium (2 mL) were seeded on glass‐bottom confocal dishes and incubated at 37 °C overnight in a humidified 5% CO₂ atmosphere. After removal of the medium, the cells were rinsed with PBS and incubated in the medium with PS‐Tz (1 µm) for 6 h. After discarding the medium and rinsing the cells with PBS twice, the cells were further incubated in the medium with or without NH₂‐Q or BCN‐Q (2 µm) for 1 h. For comparison, the cells were also incubated with the conjugate PS‐Q (1 µm) instead of PS‐Tz for 6 h without the post‐treatment or simply with BCN‐Q (2 µm) for 1 h. After being rinsed with PBS again, all the cells were replenished with Hank's Balanced Salt Solution (HBSS) (1 mL) before being examined using a Leica TCS SP8 high‐speed confocal microscope equipped with a solid‐state 638 nm laser. The photosensitizer was excited at 638 nm and its fluorescence was monitored at 650–750 nm. The images were digitized and analyzed using a Leica Application Suite X software.

Xue E.Y., Yang C., Zhou Y, & Ng D.K. (2023). A Bioorthogonal Antidote Against the Photosensitivity after Photodynamic Therapy. Advanced Science, 11(11), 2306207.

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Fluorescence Imaging of Cellular Uptake

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Approximately 2 × 10⁵ HT29 cells in RPMI medium (2 mL) or HepG2 or HEK-293 cells in DMEM medium (2 mL) were seeded on a confocal dish and incubated overnight at 37 °C with 5% CO₂. After removal of the medium, the cells were incubated with corresponding drugs. The medium was then removed and the cells were rinsed with PBS before being viewed with a Leica TCS SP8 high speed confocal microscope equipped with a solid-state 638 nm laser. The ZnPc was excited at 638 nm and monitored at 650–750 nm. The images were digitised and analysed using a Leica Application Suite X software.

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Cellular Localization of Gal-(ZnPc*)2-NP

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Senescent HeLa cells
were incubated with Gal-(ZnPc*)₂-NP (2 μM) in a serum-free medium at 37 °C
for 2 h, followed by incubation in the culture medium for 2 h, as
described above. After being rinsed with PBS twice, the cells were
stained with LysoTracker Green DND-26 (Thermo Fisher Scientific Inc.,
L7526) (2 μM), MitoTracker Green FM (Thermo Fisher Scientific
Inc., M7514) (0.2 μM), or ER-Tracker Green (Thermo Fisher Scientific
Inc., E34251) (1 μM) in a serum-free medium at 37 °C for
30, 15, or 15 min, respectively. The solutions were then removed,
and the cells were rinsed with PBS twice before being examined with
a Leica TCS SP8 high-speed confocal microscope equipped with a 488
nm laser and a 638 nm laser. All the trackers were excited at 488
nm, and their fluorescence was monitored at 500–570 nm, while ZnPc* was excited at 638 nm, and its fluorescence was monitored
at 650–750 nm. The images were digitized and analyzed using
Leica Application Suite X software.

Chu J.C., Xiong J., Wong C.T., Wang S., Tam D.Y., García-Fernández A., Martínez-Máñez R, & Ng D.K. (2023). Detection and Elimination of Senescent Cells with a Self-Assembled Senescence-Associated β-Galactosidase-Activatable Nanophotosensitizer. Journal of Medicinal Chemistry, 67(1), 234-244.

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Extrinsic Enzyme-Activated Photosensitizing System

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A mixture of 10 (61 mg, 0.04 mmol) and K2CO3 (17 mg, 0.12 mmol) in MeOH/CHCl3 (1:4 v/v, 10 mL) was stirred at room temperature for 4 h. After the consumption of 10 as indicated by TLC, the solvent was evaporated under reduced pressure, and the residue was purified by chromatography on a silica gel column using CHCl3/ MeOH (10:1 v/v) as the eluent to afford gal-DSBDP (44 mg, 80%) as a green solid. 1 h. For the study of the extrinsic enzyme-activated photosensitizing system, the cells, after being rinsed with PBS, were incubated with cRGD-β-Gal or the native β-Gal (10 unit mL -1 ) in a serum-free medium at 37 °C for 1 h. After removing the medium, the cells were rinsed with PBS twice and then treated with gal-DSBDP (4 µM) in a serum-free medium at 37 °C for 30 min. The cells were rinsed with PBS twice, followed by post-incubation in a serum-free medium for further 3 h. The solution was then removed, and the cells were rinsed with PBS twice before being examined using a Leica TCS SP8 high speed confocal microscope equipped with solid-state 488 and 638 nm lasers. The GFP was excited at 488 nm and its fluorescence was monitored at 500-530nm. The photosensitizer was excited at 638 nm and its fluorescence was monitored at 650-750 nm. The images were digitized and analyzed using a Leica Application Suite X software.

Xiong J., Chu J.C.H., Fong W.P., Wong C.T.T, & Ng D.K.P. (2022). Specific Activation of Photosensitizer with Extrinsic Enzyme for Precisive Photodynamic Therapy. Journal of the American Chemical Society, 144(23).

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