ORCA-R2 Digital Video Gallery

The Hamamatsu ORCA-R2 is the next generation scientific digital CCD camera system operating at a quantum efficiency of over 70 percent. The sensitivity of Hamamatsu's proprietary ER-150 interline CCD is approximately twice as good as the Sony ICX-285 at 700 nanometers. The rapid readout rate of the ORCA-R2 is 16.2 frames per second with low readout noise at a full resolution of 1344 x 1024 pixels. With 8 x 8 binning, a readout of 115.1 frames per second is possible. The digital videos contained in this gallery were gathered with an ORCA-R2 camera attached to an Olympus DSU spinning disk confocal microscope.

U2OS Cells (Human Bone Osteosarcoma)

Cellular products of the U2OS cell line include osteosarcoma derived growth factor (ODGF). J. Ponten and E. Saksela derived this line (originally 2T) in 1964 from a moderately differentiated sarcoma of the tibia of a 15-year-old Caucasian girl. Viruses were not detected during co-cultivation with WI-38 cells or in CF tests against SV40, RSV, or adenoviruses.

mApple fused to histone H2B and mEmerald fused to C-Src - The nucleosome is the basic repeating unit of chromatin, which enables large strands of DNA to be packed into a tiny nucleus while still allowing accessibility and functionality.

mApple fused to histone H2B and mEmerald fused to Rab5a - The digital video sequences presented in this section visualize the dynamic activity of endosomes and show their position in relation to the nucleus.

mEmerald fused to an ER targeting signal and mCherry fused to a mitochondrial targeting signal - The proximity of the endoplasmic reticulum to the mitochondrial network was visualized in the digital video sequences presented in this section.

mEmerald fused to alpha-actinin and mCherry fused to farnesyl - Due to their interaction with the microfilament actin, members of the cross-linking and bundling protein family known as alpha-actinin are popular targets for fluorescently tagging the cytoskeleton.

mCherry fused to histone H2B and mEmerald fused to C-Src - The phosphorylation of proteins by tyrosine kinases is critical for signal transduction in multicellular animals. Generally animal cells contain at least one of the Src families of tyrosine kinases.

mEmerald fused to C-Src and mCherry fused to lifeact - Microfilaments were visualized in the human bone osteosarcoma cells featured in the digital video sequences in this section with mCherry fused to lifeact.

mEmerald fused to a Golgi targeting signal and mCherry fused to an ER targeting signal peptide - Initially much of the scientific community questioned whether or not the Golgi apparatus was an actual organelle.

EGFP fused to a peroxisomal targeting signal peptide and mCherry fused to an endosomes marker - Endosomes and peroxisomes are both types of membrane-bound compartments found in eukaryotic cells.

mApple fused to histone H2B and mEmerald fused to lifeact - The ability to visualize F-actin in living cells is crucial to the success of a wide variety of scientific investigations and experiments.

Fox Lung Cells (Normal Gray Fox; FoLu Cell Line)

The FoLu line was established from the lung tissue of an adult female grey fox. The cells are susceptible to vesicular stomatitis, herpes simplex, and vaccinia viruses, but are resistant to poliovirus. FoLu cells are negative for reverse transcriptase, indicating the lack of integral retrovirus genomes.

mEmerald fused to histone H2B and mCherry fused to a mitochondrial targeting signal peptide - Mitochondria exhibit significant flexibility, and when live cells are examined it becomes clear that the organelles are almost continuously shifting shape and moving.

mEmerald fused to zyxin and mCherry fused to lifeact - In migrating cells, focal adhesions are very dynamic structures that can contain more than 600 different proteins bundled together into a complex superstructure.

mEmerald fused to lifeact and mCherry fused to zyxin - Zyxin is a zinc-binding phosphoprotein that in humans is encoded by the ZYX gene. In live cells, zyxin concentrates at focal adhesions and along the intracellular actin cytoskeletal network.

mEmerald fused to EB3 and mApple fused to zyxin - In the featured digital video sequences, the relationship between microtubules and focal adhesions, which were visualized with mApple fused to zyxin, is explored.

YPet fused to a mitochondrial targeting signal and mApple fused to a Golgi targeting signal - The Golgi apparatus receives tiny vesicles filled with proteins and lipids initially built in the endoplasmic reticulum.

EGFP fused to a Golgi targeting signal and mCherry fused to a mitochondrial targeting signal - The cis face of a Golgi stack is located near the endoplasmic reticulum and is the end where molecules enter for processing.

mCherry fused to histone H2B and mEmerald fused to C-Src - Tyrosine kinases are enzymes that are capable of transferring a phosphate group from adenosine triphosphate (ATP) to a tyrosine residue.

mEmerald fused to histone H2B and mCherry fused to farnesyl -The fluorescent protein Emerald and its monomeric form (mEmerald) were produced via the directed mutation of EGFP to generate a high-performance version.

mEmerald fused to alpha-Actinin and mCherry fused to lifeact - In the digital video sequences featured in this section, cytoskeletal actin is targeted with both mEmerald fused to alpha-actinin and a mCherry-lifeact fusion.

mEmerald fused to alpha-actinin and mCherry fused to a mitochondrial targeting signal peptide - The number of mitochondria in cells is highly variable and depends upon current cellular metabolic needs.

mEmerald fused to alpha-actinin and mCherry fused to a peroxisomal targeting signal peptide - Peroxisomes and lysosomes, which are shown in juxtaposition in these videos, differ in function and enzymatic contents.

mEmerald fused to lifeact and mApple fused to alpha-actinin - A directed evolution approach was used to produce mApple and several other red fluorescent protein variants belonging to the “mFruit” series of fluorescent proteins.

mEmerald fused lifeact and mApple fused to paxillin - Paxillin is an adaptor protein associated with focal adhesions, the close points of contact between a cell and the extracellular matrix.

mEmerald fused to a mitochondrial targeting signal and mApple fused to a Golgi targeting signal - The majority of the proteins conveyed to the endoplasmic reticulum possess a sequence of 5 to 10 hydrophobic amino acids on their N-terminal end.

mEmerald fused to a mitochondrial targeting signal and mCherry fused to a Golgi targeting signal - In the FoLu cells presented in this section, mitochondria were visualized with mEmerald, a green variant of EGFP with improved photostability and brightness.

mEmerald fused to a mitochondrial targeting signal peptide and mApple fused to histone H2B - Each mitochondrion in a cell is encircled by two specialized membranes that delineate a narrow inter-membrane space and the much bigger internal matrix.

mEmerald fused to a mitochondrial targeting signal peptide and mCherry fused to histone H2B - The energy that mitochondria create is in the form of adenosine triphosphate, or ATP, which is produced by the conversion of oxygen and nutrients.

mEmerald fused to a mitochondrial targeting signal and mCherry fused to Rab11a - Many proteins that interact with Rab11a have already been identified. One Rab11a interacting protein of particular interest is myosin Vb.

mEmerald fused to an ER targeting signal and mCherry fused to a mitochondrial targeting signal - The endoplasmic reticulum consists of an extensive interconnected network of flattened sacs and branching tubules that stretch throughout the cytoplasm.

mEmerald fused to Rab11a and mCherry fused to Rab7a - The first of the Rab proteins were identified in the mid-1980s. Since then, much has been learned about the small proteins (20-25 kiloDalton), though only about half have yet been extensively characterized.

mEmerald fused to paxillin and mApple fused to tubulin - In order for multicellular life to be possible, cells must be able to attach to the extracellular matrix. The process of attachment occurs via the construction of cell adhesion complexes.

mEmerald fused to clathrin and mApple fused to a Golgi targeting signal - The protein clathrin, which was first isolated in 1975 by British scientist Barbara Pearse, is best known for its triskelion form and its important role in coated vesicle formation.

mEmerald fused to light chain myosin and mCherry fused to lifeact - As their name suggests, myosins were initially believed to be present only in muscle cells, but members of the large superfamily of myosins can be found in almost all eukaryotic cells.

EGFP fused to vinculin - The protein vinculin is involved with focal adhesion and adherens junctions, which are complexes that operate as nucleation sites for filamentous actin and as crosslinkers between the external medium, plasma membrane, and cytoskeleton.

mEmerald fused to clathrin and mCherry fused to peroxisomes - The protein clathrin is best known for its role in coated vesicle formation and its triskelion form. Peroxisomes contain degradative enzymes, such as catalase and D-amino acid oxidase.

mEmerald fused to Golgi and mCherry fused to mitochondrial targeting signal - Many of the proteins transmitted to the endoplasmic reticulum possess a sequence of 5 to 10 hydrophobic amino acids on their N-terminal end.

Pig Kidney Cells (LLC-PK1 Cell Line)

Cellular products include plasminogen activator. The line was derived from the normal kidney of a male pig (Hampshire strain) that was 3 to 4 weeks of age.

mCherry fused to a peroxisomal targeting signal (SKL) and mEmerald fused to human histone H2B - Peroxisomes are organelles that oxygen in order to function properly, a characteristic that would have been especially important millions of years ago.

mApple fused to VE-cadherin and mEGFP fused to VE-cadherin - A variety of cellular molecules, including integrins, selectins, catenins, and cadherins, work together to facilitate interaction between cells and between cells and the extracellular matrix.

mEmerald fused to a mitochondrial targeting signal and mCherry fused to Rab7a - The digital video sequences presented in this section visualize the dynamic activity of mitochondria and endosomes.

mEmerald fused to EB3 and mCherry fused to histone H2B - Microtubules are one of the three basic components of the cytoskeleton and also function in the movement of other organelles throughout the cytoplasm.

mEmerald fused to an ER targeting signal and mApple fused to histone H2B - The region of the ER studded with ribosomes is termed the rough endoplasmic reticulum. The ribosomes are responsible for assembling amino acids into protein units.

mEmerald fused to an ER targeting signal and mCherry fused to C-Src - The rough endoplasmic reticulum is directly continuous with the nuclear envelope and is thought to have evolved simultaneously with the nucleus in early cells.

mCherry fused to Rab4a and mEmerald fused to Rab5a - Two of the Rab family of proteins involved in vesicle trafficking were targeted in a specimen of pig kidney cells. Rab5a was tagged with mEmerald and Rab4a was fluorescently labeled with mCherry.

mEmerald fused to tubulin and mCherry fused to Rab5a - The digital video sequences presented in this section visualize the dynamic activity of endosomes, which were targeted with mCherry fused to the early endosomal marker Rab5a.

mEmerald fused to VE-cadherin - Research involving VE-cadherin and inter-endothelial cell junctions suggests that the protein is significantly involved in the maintenance of proper endothelial permeability.

mEGFP fused to Cx43 - Connexins are a family of proteins present in vertebrates that form intercellular channels in the plasma membranes of adjacent cells. These gap junctions facilitate free passage of ions and various small molecules between the cells.

HeLa Cells (Human Cervical Carcinoma)

Cellular products of HeLa cells are keratin and lysophosphatidylcholine (lyso-PC), which induces AP-1 activity and c-jun N-terminal kinase activity (JNK1) by a protein kinase C-independent pathway. The cells are positive for keratin by immunoperoxidase staining. HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences. P53 expression was reported to be low, and normal levels of pRB (retinoblastoma suppressor) were found. HeLa cells have been used to screen for Escherichia coli strains with invasive potential and are useful for transfections. The cells are susceptible to Adenovirus type 3, Encephalomyocarditis virus, Poliovirus 1, Poliovirus 2, and Poliovirus 3, but are negative for reverse transcriptase, indicating the lack of integral retrovirus genomes.

mApple fused to histone H2B and mEmerald fused to C-Src - Located in the nuclei of eukaryotic cells, histones are closely associated with DNA. The proteins, all but one of which is comprised of 100 to 150 residues, function in the formation of nucleosomes.

mCherry fused to histone H2B and mEmerald fused to C-Src - For many years after they were initially identified, histones were widely believed to primarily play a supportive role, acting as a type of packing material around which DNA was wound.

Rabbit Kidney Cells (RK-13 Cell Line)

Rabbit kidney cells are positive for keratin by immunoperoxidase staining and are positive for bovine viral diarrhea virus (BVDV). The RK-13 line, which was derived from the kidney of a 5-week-old rabbit, is susceptible to B virus, herpes simplex, pseudorabies virus, vaccinia, rabbitpox, myxoma, simian adenoviruses, and rubellavirus.

mApple fused to histone H2B and mEmerald fused to C-Src - The C-Src gene is closely related to the viral-sarcoma gene (V-SRC) present in certain cancer-causing retroviruses. The normal cellular gene, C-Src, was first discovered in chickens in the late 1970s.

mEmerald fused to zyxin and mCherry fused to lifeact - In live cells, zyxin, a zinc-binding phosphoprotein that in humans is encoded by the ZYX gene, concentrates along the intracellular actin cytoskeletal network and at focal adhesions.

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