Dental pulp stem cell (DPSC) were grown and differentiated on 3-well chamber slides (Ibidi, Planegg, Germany) coated with poly-D-lysine. Cells were fixed using a 4% paraformaldehyde solution for 10 min. Once fixed, cells were blocked and permeabilized using PBS with 1% BSA, 10% FBS, and 0.3% Triton X-100 for 1 h. Primary antibodies were diluted to 1:500 for anti-Beta Tubulin (Millipore, ab9354) and anti-TOMM20 (Santa Cruz, sc-17764) in the blocking solution and incubated overnight at 2–8°C with agitation. After overnight incubation, the slides were washed 3x with PBS-T for 10 min before the secondary antibodies, Goat anti-mouse Alexa Fluor 488 (Life Technologies, A11029) and Goat anti-chicken Alexa Fluor 594 (Life Technologies, A11042) were added at a 1:1000 dilution. Slides were incubated at room temperature for 1 h and then washed again 3× for 10 min each. Finally, Prolong Gold Antifade with DAPI (Fisher Scientific, Waltham, MA) was applied for mounting. Slides were imaged on a Zeiss 710 confocal microscope at 63× magnification using Z-stacking to image the entire neuron via ZEN software (Black Edition).
3 well chamber slides
Manufactured by Ibidi
Sourced in Germany
The 3-well chamber slides are a type of laboratory equipment designed for cell culture applications. Each slide features three separate wells, providing a convenient platform for conducting multiple experiments or observations simultaneously. The slides are made of high-quality materials and are suitable for a variety of cell types and experimental setups.
Automatically generated - may contain errors
Lab products found in correlation
710 confocal microscope, by Zeiss (6 mentions)
Prolong gold antifade with dapi, by Thermo Fisher Scientific (2 mentions)
Anti β tubulin, by Merck Group (1 mentions)
Goat anti chicken alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Anti tomm20, by Santa Cruz Biotechnology (1 mentions)
2 protocols using 3 well chamber slides
1
Immunostaining of Dental Pulp Stem Cells
2
Immunocytochemistry of Differentiated DPSC
Check if the same lab product or an alternative is used in the 5 most similar protocols
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DPSC were grown and differentiated on 3-well chamber slides (Ibidi, Planegg, Germany) coated with poly-D-lysine. Cells were fixed using a 4% paraformaldehyde solution for 10 minutes. Once fixed, cells were blocked and permeabilized using PBS with 1% BSA, 10% FBS, and 0.3% Triton X-100 for 1 hour.
Primary antibodies were diluted to 1:500 for a-Beta Tubulin (Millipore, ab9354) and a-TOMM20 (Santa Cruz, sc-17764) in the blocking solution and incubated overnight at 2-8 °C with agitation. After overnight incubation, the slides were washed 3x with PBS-T for 10 minutes before the secondary antibodies, Goat a-mouse Alexa Fluor 488 (Life Technologies, A11029) and Goat a-chicken Alexa Fluor 594 (Life Technologies, A11042) were added at a 1:1000 dilution. Slides were incubated at room temperature for 1 hour and then washed again 3x for 10 minutes each. Finally, Prolong Gold Antifade with DAPI (Fisher Scientific, Waltham, MA) was applied for mounting. Slides were imaged on a Zeiss 710 confocal microscope at 63x magnification using Z-stacking to image the entire neuron via ZEN software (Black Edition).
Primary antibodies were diluted to 1:500 for a-Beta Tubulin (Millipore, ab9354) and a-TOMM20 (Santa Cruz, sc-17764) in the blocking solution and incubated overnight at 2-8 °C with agitation. After overnight incubation, the slides were washed 3x with PBS-T for 10 minutes before the secondary antibodies, Goat a-mouse Alexa Fluor 488 (Life Technologies, A11029) and Goat a-chicken Alexa Fluor 594 (Life Technologies, A11042) were added at a 1:1000 dilution. Slides were incubated at room temperature for 1 hour and then washed again 3x for 10 minutes each. Finally, Prolong Gold Antifade with DAPI (Fisher Scientific, Waltham, MA) was applied for mounting. Slides were imaged on a Zeiss 710 confocal microscope at 63x magnification using Z-stacking to image the entire neuron via ZEN software (Black Edition).
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