Live-cell imaging was performed on an upright Olympus microscope (BX51WI) coupled to an EM-CCD camera (Ixon, Andor). Cells were imaged with a water-immersion 60× objective (Olympus). Excitation was provided by an X-cite 120Q light source (Lumen Dynamics). Appropriate filters were used (in nm): Excitation: 470/40; Emission: 525/50; Dichroic: 495, long pass. The image pixel scale was calculated by dividing the camera pixel size (16 μm) by the lens magnification (60×) yielding a pixel size of 0.27 μm. Before constant perfusion with a Cole–Parmer Master-Flex pump (∼4 ml/min), aCSF (126 mM NaCl, 24 mM NaHCO3, 10 mM d -Glucose, 2.5 mM KCL, 2 mM CaCl2, 1 mM MgCl, 1 mM NaH2PO4, 5 mM Sodium Pyruvate) was pre-equilibrated for 20 min with 95% O2 and 5% CO2 to establish a pH of 7.4. Temperature of the waterbath was constantly measured using a digital Thermometer (Hanna Instruments) and maintained at 37 °C. Any focus drift was corrected manually. Protocols were adapted to achieve minimal bleaching conditions. Imaging of SEP-tagged GABAARs was done for 60 min at a rate of one frame every 20 s (180 frames, 48.8 ms exposure, no averaging). For imaging of intracellular Ca2+ using fluo4 (1 μM, Molecular Probes, Invitrogen) hippocampal neurons were incubated for 30 min at 37 °C. After washing twice, fluo4-imaging was done for 60 min (720 frames, 5 ms exposure, no averaging) at 1 frame every 5 s.
X cite 120q light source
Manufactured by Excelitas
Sourced in Canada, United States
The X-Cite 120Q is a light source designed for use in a variety of laboratory applications. It provides stable, uniform illumination across a wide range of wavelengths. The X-Cite 120Q is a compact and easy-to-use device that offers consistent performance for various microscopy and imaging techniques.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision 4, by Zeiss (2 mentions)
Apotome, by Zeiss (2 mentions)
Axiocam mrc, by Zeiss (2 mentions)
Digital thermometer, by Hanna Instruments (1 mentions)
Bx51wi, by Oxford Instruments (1 mentions)
Water immersion 60 objective, by Olympus (1 mentions)
Fluo 4, by Thermo Fisher Scientific (1 mentions)
Emccd camera, by Oxford Instruments (1 mentions)
Axiovert 40 cfl microscope, by Zeiss (1 mentions)
Zen 2011, by Zeiss (1 mentions)
Lumar v12, by Zeiss (1 mentions)
Axiocam mrm, by Zeiss (1 mentions)
Axio imager m2, by Zeiss (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
5 protocols using x cite 120q light source
1
Live-cell Imaging of GABA Receptors and Calcium Signaling
2
Fluorescent Microscopy of Transfected Cells
3
Postmortem Brain Analysis of A-T Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen postmortem brain samples of diseased A-T patients and age-matched controls were requested from the NeuroBioBank, National Institutes of Health (NIH). All samples were thoroughly anonymized prior distribution. The US Human Studies Board characterizes these tissues as exempt. All studies conducted using human tissue strictly complied with the ethical standard of NIH and the Ethics Committee on Research Practices of the HKUST. Frozen sections were prepared from brain blocks and were used for immunohistochemistry purposes. All tissue sections were examined and imaged on a fluorescent microscope (Olympus BX53 with DP80 camera) equipped with 20× (UPlanSApo; 0.75 NA) objective, using an X-Cite 120Q light source (Excelitas Technologies Corp.).
4
Imaging Larval Pelts and Fixed Embryos
5
Perfused Brain Tissue Processing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After data acquisition was completed, rats were deeply anesthetized with 3% isoflurane and urethane (i.p. 10 mL/kg 20%). Next, rats were perfused transcardially with 0.9% NaCl followed by 4% paraformaldehyde (PFA). Brains were extracted and stored overnight in 4% PFA in phosphate buffer (PB) at 4°C after which they were transferred to 0.05 M PB for further processing.
Brains were sliced into 100 or 50 μm coronal sections with a vibratome, rinsed with PB (0.05 BM) and mounted using a Mowiol solution (Clairant GmbH, Frankfurt am Main, Germany; Narayanan et al., 2014 (link)). Probe placement was verified visually using an Olympus BX51 microscope with a 4× air objective or 40× oil objective. For chronic recordings, we used the electrode tract to retrieve probe placement. For acute recordings, we dipped the probe in DiI (Thermo Fisher Scientific, Waltham, MA, USA) and used a X-Cite 120 Q light-source (Excelitas Technologies Corp., Waltham, MA, USA), to visualize the electrode tract.
Brains were sliced into 100 or 50 μm coronal sections with a vibratome, rinsed with PB (0.05 BM) and mounted using a Mowiol solution (Clairant GmbH, Frankfurt am Main, Germany; Narayanan et al., 2014 (link)). Probe placement was verified visually using an Olympus BX51 microscope with a 4× air objective or 40× oil objective. For chronic recordings, we used the electrode tract to retrieve probe placement. For acute recordings, we dipped the probe in DiI (Thermo Fisher Scientific, Waltham, MA, USA) and used a X-Cite 120 Q light-source (Excelitas Technologies Corp., Waltham, MA, USA), to visualize the electrode tract.
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!