Epifluorescence excitation was by light-emitting diode in a dual–lamp house controlled by an OptoLED system (Cairn Research) at 470-/40-nm (and 572-/35-nm; Fig. 1, A and B ) excitation; fluorescence emitted at 520/40 nm (and 630/60 nm; Fig. 1, A and B ) was detected with an electron-multiplying CCD camera (Andor Technology; filters from Chroma Technology Corp.). The second channel of the lamp house was used to connect the optic fiber coming from the photolysis laser. After each experiment, a series of epifluorescence images was taken to perform morphological reconstructions and to measure the distance between the release sites and the soma. ImageJ (National Institutes of Health) was used to make the stacks and composite images.
Optoled system
Manufactured by Cairn Research
Sourced in United Kingdom
The OptoLED system is a high-performance LED illumination solution designed for various research and experimental applications. The core function of the OptoLED system is to provide precise and programmable light control, enabling researchers to accurately manipulate light parameters as required for their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Axioskop 2 fs microscope, by Zeiss (1 mentions)
Bx51wi compound microscope, by Olympus (1 mentions)
Multiclamp 700b amplifier, by Molecular Devices (1 mentions)
Digidata 1440a, by Molecular Devices (1 mentions)
Clampfit 10, by Molecular Devices (1 mentions)
Fluo 4ff, by Thermo Fisher Scientific (1 mentions)
Vt1200 vibratome, by Leica (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
4 protocols using optoled system
1
Epifluorescence Imaging Reconstruction
2
In Vitro Acute Hippocampal Slice Electrophysiology
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preparation and maintenance of acute slices in vitro and composition of extracellular and intracellular solutions for electrophysiological experiments were performed according to previously published procedures (Mańko et al., 2012 (link)) and as described in Alcami et al. (2012) (link), with minor modifications. Optical stimulation of hippocampal ChR2-expressing afferents in the BA in vitro was performed using an optoLED system (Cairn Research) mounted on a Zeiss Axioskop 2 FS microscope. The spot size corresponded to an area of about 200 μm. Optical TBS consisted of ten trains delivered at the same frequency as for the in vivo experiments. Analysis of electrophysiological signals was performed using MATLAB custom scripts. Full details on ex vivo electrophysiology, optogenetics, and analysis are given in the Supplemental Information .
3
Electrophysiological Recording of Larval Motoneurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CNS was removed from wall climbing third instar larvae and placed dorsal surface up on a sylgard-coated coverslip (Dow Corning, MI, USA). Saline was composed of (in mM): 135 NaCl, 5 KCl, 4 MgCl2, 2 CaCl2, 5 TES, and 36 sucrose, pH 7.15 with NaOH. The ventral nerve cord (VNC) which contains motoneurons was viewed using an Olympus BX51-WI compound microscope with a 20x water-immersion lens. For electrophysiology, the neurolemma (glial sheath) covering the VNC was removed as described in [16 (link)] to gain access to the aCC motoneuron. Loose-patch recordings were obtained using a MultiClamp 700B amplifier and Digidata 1440A (Molecular Devices, Sunnyvale, CA). Borosilicate glass capillaries were used to pull recording electrodes (unpolished) with resistances between 3–5 MΩ. Data were acquired with a sampling rate of 20 kHz, filtered with a low-pass filter of 10 kHz and analysed in Clampfit 10.4 (Molecular Devices). Excitation of GCaMP was achieved using an OptoLED system (Cairn Research) with a 470 nm LED. The frame rate of acquisition was 5/s and frame duration of 200 ms with a QImaging Exi aqua camera (Photometrics, UK).
4
Fluorescent Calcium Imaging in Brain Slices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For all comparative tests on Fluo4FF (Thermo Fisher Scientific, Waltham, MA, USA), Cal520FF (AAT Bioquest, Sunnyvale, CA, USA) and OG5N (Thermo Fisher Scientific), we used a set-up originally developed for fast Ca 2+ imaging [3] (link). The system was mounted on an Olympus BX51 microscope equipped for electrophysiology in brain slices and with a 60X NA=1 objective. Fluorescence was excited at 470 nm using an OptoLED system (CAIRN Research Ltd., Faversham, UK) and recorded at 530 ± 21 nm with a NeuroCCD-SMQ camera (RedShirtimaging, Decatur, GA, USA). Each indicator was purchased in its (potassium) salt form and dissolved in an aqueous solution. Physiological tests were performed in somatosensory neocortical layer-5 (L5) pyramidal neurons from brain slices prepared as described in a recent publication [12] (link). Briefly, animal manipulation was ethically carried out in accordance with European Directives 2010/63/UE on the care, welfare and treatment of animals and procedures were reviewed by the ethics committee affiliated to the animal facility of the university (D3842110001). 21-35 postnatal days old mice (C57BL/6j) were anesthetised by isofluorane inhalation and brain slices (350 µm thick) with an orientation of 15 degrees from the coronal plane were prepared using a Leica VT1200 vibratome (Leica, Wetzlar, Germany).
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!