Orca r2 monochrome camera

Five thousand SK-N-SH cells were seeded per well in 12-well plates and chemically treated the next day. After treatment, phase-contrast images were recorded (ten fields of view per well) with an Olympus IX71 inverted microscope and a Hamamatsu Orca R2 monochrome camera and randomised for blind analysis. The length of the neurites in each image was measured using the NeuroJ plugin on ImageJ⁷⁶ . Only neurites longer than the cell body were measured and an average neurite length per field was calculated.

Studies were carried out using ZF models. Fertilised eggs were incubated and WT embryos were collected at 24, 48 and 72 hpf. Antisense morpholinos sequences designed to block translation and thereby reduce Brn-3a and Brn-3b protein expression were as follows: Brn-3b- 5′-AGACATCATCATCATATTTGCGACC-3′ Brn-3a - 5′-AG CGTCTCATCCAGACTGGCGAAGA-3′. Standard Control oligo was used as a non-specific control. All morpholinos were obtained from Gene Tools, LLC (www.gene-tools.com). During preliminary studies, 1 ng, 2 ng and 4 ng of the control, Brn-3a and Brn-3b (mix) morpholinos were injected into fertilised embryos and effects on embryonic viability were analysed at different times (24, 48, 72 h) and the morphological and functional effects on the heart were analysed using live image capture, for example, Zeiss Axiovert 135 live imaging scope (Jena, Germany) with motorised stage Hamamatsu Orca R2 monochrome camera (Hamamatsu, Japan).

Whole-mount images were taken using a Leica MZFLIII microscope (Leica Microsystems, Wetzlar, Germany) fitted with a QImaging MicroPublisher 5.0 RTV camera and QCapture Pro 6.0 software (QImaging, Surrey, BC, Canada); a Zeiss AxioSkop2 microscope fitted with a Zeiss AxioCam HRc camera and Zeiss AxioVision Rel. 4.8 software (Carl Zeiss, Oberkochen, Germany); an Olympus MVX10 microscope (Olympus Corporation, Tokyo, Japan) fitted with a Zeiss AxioCam HRc camera and Zeiss AxioVision Rel. 4.8 software; an Olympus 1 × 71 inverted microscope fitted with a Hamamatsu ORCA-R2 monochrome camera and HCImage software (Hamamatsu Photonics, Hamamatsu, Japan); a Zeiss LSM 710 confocal microscope with Zeiss ZEN software; and a Zeiss 710 confocal microscope with Zeiss LSM Image Browser (version 4.2.0.121) software, which was used to create three-dimensional images and stack movies. Images of sections were taken using a Zeiss AxioSkop 2 MOT microscope fitted with a QImaging Retiga 2000R camera, a Qimaging RGB pancake and QCapture Pro 6.0 software; a Zeiss Scope.A1 microscope fitted with a Zeiss AxioCam MRm camera and Zeiss ZEN 2012 (blue edition) software; and a Zeiss LSM 780 confocal microscope with Zeiss ZEN 2011 (black edition) software. All images were further processed in Photoshop CS4 (Adobe Systems Inc., San Jose, CA) and/or ImageJ 1.50i software (NIH, Bethesda, MD).