SH-SY5Y cells were seeded on a glass coverslip in a six-well plate at a density of 40 × 103 cells per well. After 24 h, 600 µL of various samples (Aβ42 ADDLs, Aβ42 + PrP107–120, Aβ42 ADDLs + PrP107–120 and PrP107–120, all pre-dissolved and pre-incubated in 2% (v/v) DMSO and F-12 Ham medium, as described above) were added to cells for 1 h at 37 °C. Final concentrations of Aβ42 and PrP107-120 in DMEM were 3 and 0.75 µM (m.e.), respectively. After washing with phosphate buffered saline (PBS), cells were loaded with a 4-µM Fluo-4 AM probe (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA) for 10 min at 37 °C. Imaging was performed after excitation at 488 nm with a TCS SP8 confocal scanning microscopy system (Leica Microsystems, Mannheim, Germany), using a Leica Plan Apo 63x oil immersion objective, taking a series of 1-µm-thick optical sections (1024 × 1024) through the cell depth for each sample and projecting them as a single composite image by superimposition. A minimum of four images were captured for each sample and four replicates were used for each condition. Images were analyzed using Image J software (NIH, Bethesda, MD, USA).
Plan apo 63x oil immersion objective
Manufactured by Leica
Sourced in Germany
The Plan Apo 63x oil immersion objective is a high-performance microscope lens designed for advanced microscopy applications. It features a high numerical aperture and a flat image field, delivering exceptional optical performance and resolution.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using plan apo 63x oil immersion objective
1
Aβ42 and PrP107-120 Interaction Effects on SH-SY5Y Cells
2
Mitochondrial Network Labeling in HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Labeling of mitochondrial network with Mitotracker was performed by incubating HeLa cells seeded in glass coverslips with a Mitotracker red 1 µM in Opti-MEM TM (ThermoFisher) for 30 min at 37 o C and 5% CO 2 . Cells fixation was carried out by incubation with 4% PFA/PBS for 30 min at room temperature. Coverslips were then washed in PBS and incubated with 50 mM NH4Cl/PBS for 15 min at room temperature.
For immunofluorescence, cells were incubated with the primary antibodies diluted in blocking solution (1% BSA / 0.1% Saponin in PBS) for 1 hr at room temperature.
Coverslips were thereafter incubated with fluorescently-labeled secondary antibodies
(1:500 in BB) for 1hr at room temperature. To label LDs, cells were incubated with 10 µM LTox in 1xPBS for 30 minutes at room temperature. After washing with PBS, coverslips were mounted with Vectashield (Vectro Laboratories) on microscopy slides.
Images were acquired on a confocal inverted microscope SP8-X (DMI 6000 Leica).
Optical sections were acquired with a Plan Apo 63x oil immersion objective (N.A. 1.4, Leica) using the LAS-X software. Fluorescence was excited using either a 405nm laser diode or a white light laser, and later collected after adjusting the spectral windows with GaAsP PMTs or Hybrid detectors. Images from a mid-focal plane are shown.
For immunofluorescence, cells were incubated with the primary antibodies diluted in blocking solution (1% BSA / 0.1% Saponin in PBS) for 1 hr at room temperature.
Coverslips were thereafter incubated with fluorescently-labeled secondary antibodies
(1:500 in BB) for 1hr at room temperature. To label LDs, cells were incubated with 10 µM LTox in 1xPBS for 30 minutes at room temperature. After washing with PBS, coverslips were mounted with Vectashield (Vectro Laboratories) on microscopy slides.
Images were acquired on a confocal inverted microscope SP8-X (DMI 6000 Leica).
Optical sections were acquired with a Plan Apo 63x oil immersion objective (N.A. 1.4, Leica) using the LAS-X software. Fluorescence was excited using either a 405nm laser diode or a white light laser, and later collected after adjusting the spectral windows with GaAsP PMTs or Hybrid detectors. Images from a mid-focal plane are shown.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!