The exciter filter, dichroic mirror, and barrier filter can be assembled together into a component known as the filter cube. Different filter cubes can be changed during specimen viewing to change the excitation wavelength, and a series of diaphrams can be used to modify the intensity of excitation.

What is fluorescence microscopyused for

It depends, says cat behavior expert Samantha Bell. With a few simple tweaks, playing with a laser pointer doesn't need to end in anxious feelings for your cat. Here's Bell's breakdown of why laser pointers can be bad for cats and how to make laser pointer play fun and enriching. Plus, fear-free certified experts share their favorite methods of play with cats in place of the never-ending red-light saga.

This light is reflected toward the sample by a special mirror called a dichroic mirror, which is designed to reflect light only at the excitation wavelength. The reflected light passes through the objective where it is focused onto the fluorescent specimen. The emissions from the specimen are in turn, passed back up through the objective – where magnification of the image occurs –and now through the dichroic mirror.

Next, place your sample on the stage and secure it in place. Then, turn on the white light source of your microscope. Focus on your sample using the lowest powered objective by adjusting the coarse and fine focus knobs. Then, use the stage adjustment knobs to find your area of interest.

Next, place your sample on the stage and secure it in place. Then, turn on the white light source of your microscope. Focus on your sample using the lowest powered objective by adjusting the coarse and fine focus knobs. Then, use the stage adjustment knobs to find your area of interest.

Many different types of experiments can make use of fluorescent microscopy and involve different types of fluorophores One of the most common applications of fluorescent microscopy is the imaging of proteins that have been labeled with antibodies that are attached to, or “conjugated” to fluorescent compounds.. Here, an antibody towards leptospiral surface proteins was detected using a secondary antibody conjugated to alexafluor-488, which fluoresces green when excited.

Every frisky feline loves playing with her laser pointer, right? Not always. Laser pointers can be a good option for kitty cardio and human laughs, but they might leave your cat feeling frustrated and wanting to pounce on the next best thing (like your feet). This has us wondering, "Are laser pointers bad for cats?"

When it comes to performing fluorescence microscopy, the fluorophore can be just as important as the microscope itself, and the type of fluorophore being imaged dictates the excitation wavelength used and emission wavelength that’s detected. The excitation wavelengths contain a small range of energies that can be absorbed by the fluorophore and cause it to transition into an excited state. Once excited, a wide range of emissions, or transitions back to the lower energy state, are possible resulting in an emission spectrum.

Another way to highlight a specific feature with fluorescence is to integrate the code for a fluorescent protein such as green fluorescent protein, or GFP, into the DNA of an organism. The gene for GFP was originally isolated from jellyfish and can be expressed, or produced, by cultured cells in response to specific triggers or as part of a specific cell type like the tumor cells shown glowing in this image

It seems that cat experts agree—wand time is the best method of play when it comes to replicating prey. "My absolute favorite wand toy that most cats love is Da Bird," says Shannen McNee, CCBC from the Toronto Humane Society. "It makes a really cool fluttering sound and movement when you 'fly' it around in the air that your cat will go crazy for!"

Finally, make fine focus adjustments and direct the output light to the imaging camera. You will likely need to make adjustments to the exposure time for each different fluorophore or fluorescent dye used. However, it is important to keep the exposure time constant when comparing features with the same dye on different samples.

An advanced technique known as Fluorescence recovery after photobleaching, or FRAP, is performed by intentionally photobleaching a small region of a sample in order to monitor the diffusion rate of fluorescently labeled molecules back into the photobleached region.

This light is filtered by the barrier filter, which selects for the emission wavelength and filters out contaminating light from the arc lamp or other sources that are reflected off of the microscope components. Finally, the filtered fluorescent emission is sent to a detector where the image can be digitized, or it’s transmitted to the eyepiece for optical viewing.

The main components of the fluorescent microscope overlap greatly with the traditional light microscope. However the 2 main differences are the type of light source and the use of the specialized filter elements.

Janelle is a cat mum to two resident adventure kitties, Lyra and Atlas, and numerous cat and kitten fosters. Janelle and her furry family enjoy filling their days with hiking, kayaking, and seeking out the best cat-friendly destinations around the Pacific Northwest. You can follow Janelle, her adventure kitties, and adoptable fosters at @paws_pdx.

Fluorescence microscopyprinciple

The main components of the fluorescent microscope overlap greatly with the traditional light microscope. However the 2 main differences are the type of light source and the use of the specialized filter elements.

Fluorescence microscopyppt

The principle behind fluorescence microscopy is simple. As light leaves the arc lamp it is directed through an exciter filter, which selects the excitation wavelength.

What is fluorescence microscopyprocedure

Fluorescence is a phenomenon that takes place when a substance absorbs light at a given wavelength and emits light at another wavelength. Fluorescence occurs as an electron, which has been excited to a higher, and more unstable energy state, relaxes to its ground state and gives off a photon of light. The light that is responsible for excitation, or moving the electron to a higher energy state, is of shorter wavelength and higher energy than the fluorescence emission, which has a longer wavelength, lower energy, and different color.

In this video we learned about the concept of fluorescence, how fluorescence microscopy differs from light microscopy, and how to take a fluorescence image through the scope. We also learned about some basic and advanced applications that use fluorescence. Thanks for watching and don’t forget while photobleaching looks great on your teeth it’s not so good for your samples.

To begin fluorescence imaging, turn on the xenon or mercury light source and allow it to warm up for as long as 15 minutes in order for it to reach constant illumination.

Another application of fluorescence imaging is Fluorescence Speckle Microscopy which is a technology that uses fluorescently labeled macromolecular assemblies such as the F-actin network seen here, to study movement and turnover kinetics of this important cytoskeletal protein.

Advantages offluorescence microscopy

DeVoss is a fan of motion-activated toys for solo play when the humans are away—and joins the thousands of reviewers who say their cats are obsessed with the popular Potaroma Flopping Fish Cat Toy. When choosing a motion-activated toy for your cat, she says, look for one that allows your cat to catch and bite it.

Fluorescence microscopy requires a very powerful light source such as a xenon or mercury arch lamp like the one shown here. The light emitted from the mercury arc lamp is 10-100 times brighter than most incandescent lamps and provides light in a wide range of wavelengths, from ultra-violet to the infrared. This high-powered light source is the most dangerous part of the fluorescence microscope setup as looking directly into unfiltered light can seriously damage your retinas and mishandling the bulbs can cause them to explode.

Amy Shojai has championed cats and dogs for over 40 years. As one of the best known pet journalists, her career focuses on empowering pet lovers to make the best informed choices for their cats and dogs. She enjoys translating complicated "medicalese" into information everyone easily understands, to improve the lives of cats and dogs we love.

An advanced technique known as Fluorescence recovery after photobleaching, or FRAP, is performed by intentionally photobleaching a small region of a sample in order to monitor the diffusion rate of fluorescently labeled molecules back into the photobleached region.

The principle behind fluorescence microscopy is simple. As light leaves the arc lamp it is directed through an exciter filter, which selects the excitation wavelength.

Fluorescence microscopy requires a very powerful light source such as a xenon or mercury arch lamp like the one shown here. The light emitted from the mercury arc lamp is 10-100 times brighter than most incandescent lamps and provides light in a wide range of wavelengths, from ultra-violet to the infrared. This high-powered light source is the most dangerous part of the fluorescence microscope setup as looking directly into unfiltered light can seriously damage your retinas and mishandling the bulbs can cause them to explode.

Whichever mode of play you and your cat choose, experts recommend following playtime with mealtime. "This helps complete the predatory cycle of 'hunt, catch, kill, eat' and helps them feel as satisfied as they can be", Bell says.

Finally, make fine focus adjustments and direct the output light to the imaging camera. You will likely need to make adjustments to the exposure time for each different fluorophore or fluorescent dye used. However, it is important to keep the exposure time constant when comparing features with the same dye on different samples.

"Cats feel their happiest and most confident when they feel like predators," Bell explains. So, yes, they can love the thrill of hunting the laser pointer light. But, as we all know, your cat can never actually capture the red dot and fun could quickly become frustration.

Use offluorescencemicroscope in microbiology

Fluorescence is a phenomenon that takes place when a substance absorbs light at a given wavelength and emits light at another wavelength. Fluorescence occurs as an electron, which has been excited to a higher, and more unstable energy state, relaxes to its ground state and gives off a photon of light. The light that is responsible for excitation, or moving the electron to a higher energy state, is of shorter wavelength and higher energy than the fluorescence emission, which has a longer wavelength, lower energy, and different color.

This light is filtered by the barrier filter, which selects for the emission wavelength and filters out contaminating light from the arc lamp or other sources that are reflected off of the microscope components. Finally, the filtered fluorescent emission is sent to a detector where the image can be digitized, or it’s transmitted to the eyepiece for optical viewing.

Because your cat can't catch and "kill" the laser pointer light, she can't complete the predatory sequence. This, Bell says, could leave your cat feeling unsatisfied. "If a cat is riled up in predator mode, has been "hunting" the laser, and isn't provided something appropriate to attack or bite, they may attack the closest thing to them that moves—like you or another pet."

But, she adds, this doesn't mean you need to toss your cat's favorite laser pointer in the trash. "If you always follow laser pointer time with wand toy time, your cat will be able to physically catch the 'prey' they've been hunting and feel satisfied," Bell says. "Another little trick I love is aiming the laser at a treat at the end of wand toy playtime. Then they can eat their 'prey!'"

Fluorescencemicroscope diagram

Fluorescence microscopy is a very powerful analytical tool that combines the magnifying properties of light microscopy with visualization of fluorescence. Fluorescence is a phenomenon that involves absorbance and emission of a small range of light wavelengths by a fluorescent molecule known as a fluorophore. Fluorescence microscopy is accomplished in conjunction with the basic light microscope by the addition of a powerful light source, specialized filters, and a means of fluorescently labeling a sample. This video describes the basic principles behind fluorescence microscopy including the mechanism of fluorescence, the Stoke’s shift, and photobleaching. It also gives examples of the numerous ways to fluorescently label a sample including the use of fluorescently tagged antibodies and proteins, nucleic acid fluorescent dyes with, and the addition of naturally fluorescent proteins to a specimen. The major components of the fluorescence microscope including a xenon or mercury light source, light filters, the dichroic mirror, and use of the shutter to illuminate the sample are all described. Finally, examples of some of the many applications for fluorescence microscopy are shown.

When it comes to performing fluorescence microscopy, the fluorophore can be just as important as the microscope itself, and the type of fluorophore being imaged dictates the excitation wavelength used and emission wavelength that’s detected. The excitation wavelengths contain a small range of energies that can be absorbed by the fluorophore and cause it to transition into an excited state. Once excited, a wide range of emissions, or transitions back to the lower energy state, are possible resulting in an emission spectrum.

Fluorescence microscopy combines the magnifying properties of the light microscope with fluorescence technology that allows the excitation of- and detection of emissions from- fluorophores – fluorescent chemical compounds. With fluorescence microscopy, scientists can observe the location of specific cell types within tissues or molecules within cells.

Cats have an inherent predatory drive, and the little red light has just the stuff to ramp up their feisty instincts. Lasers can slither up walls, rapidly change speed, and zigzag every which way. It looks like living prey to your huntress and kickstarts your cat's predatory sequence (stalk, capture, kill, eat).

Molly DeVoss, CFTBS, CCBC, FFCT, CRM, is the founder of Cat Behavior Solutions and her cat Pico de Gato doesn't always need a toy to enjoy play. "My cat and I play hide-and-seek! He follows me around the house and when I go into another room, I look back at him, make eye contact, then dart behind the wall or door," DeVoss shares. "He always knows where I am, but he likes to jump out and 'get' me." To complete the predatory sequence, reward your avid hunter with a treat once he finds his prey—aka you!

To begin fluorescence imaging, turn on the xenon or mercury light source and allow it to warm up for as long as 15 minutes in order for it to reach constant illumination.

The difference between the peak of the absorption, or excitation curve and the peak of the emission curve is known as Stoke’s Shift. The greater the distance in this shift, the easier it is to separate the two different wavelengths. Additionally, any overlapping spectrum needs to be removed by the components of the filter cube for reduced background and improved image quality.

The exciter filter, dichroic mirror, and barrier filter can be assembled together into a component known as the filter cube. Different filter cubes can be changed during specimen viewing to change the excitation wavelength, and a series of diaphrams can be used to modify the intensity of excitation.

Ready to ditch the laser and try something new with your feline? We asked fear-free certified cat behavior experts to share their tried-and-true ways to play with cats in place of the laser pointer.

What is fluorescence microscopyin microbiology

Another way to highlight a specific feature with fluorescence is to integrate the code for a fluorescent protein such as green fluorescent protein, or GFP, into the DNA of an organism. The gene for GFP was originally isolated from jellyfish and can be expressed, or produced, by cultured cells in response to specific triggers or as part of a specific cell type like the tumor cells shown glowing in this image

Exposure of the fluorophore to prolonged excitation will cause it to photobleach, which is a weakening or loss of fluorescence. To reduce photobleaching, you can add an anti-fade mounting medium to the slide and seal the edges with nail polish. The slide should also be kept in the dark when not being imaged.

This light is reflected toward the sample by a special mirror called a dichroic mirror, which is designed to reflect light only at the excitation wavelength. The reflected light passes through the objective where it is focused onto the fluorescent specimen. The emissions from the specimen are in turn, passed back up through the objective – where magnification of the image occurs –and now through the dichroic mirror.

Another application of fluorescence imaging is Fluorescence Speckle Microscopy which is a technology that uses fluorescently labeled macromolecular assemblies such as the F-actin network seen here, to study movement and turnover kinetics of this important cytoskeletal protein.

In this video we learned about the concept of fluorescence, how fluorescence microscopy differs from light microscopy, and how to take a fluorescence image through the scope. We also learned about some basic and advanced applications that use fluorescence. Thanks for watching and don’t forget while photobleaching looks great on your teeth it’s not so good for your samples.

If you're not sure if your cat has become frustrated when playing with her laser pointer, you're not alone. "Cats are quite subtle with their body language and sometimes play mode body language looks quite similar to annoyed body language," Bell says.

Many different types of experiments can make use of fluorescent microscopy and involve different types of fluorophores One of the most common applications of fluorescent microscopy is the imaging of proteins that have been labeled with antibodies that are attached to, or “conjugated” to fluorescent compounds.. Here, an antibody towards leptospiral surface proteins was detected using a secondary antibody conjugated to alexafluor-488, which fluoresces green when excited.

Fluorescence microscopy combines the magnifying properties of the light microscope with fluorescence technology that allows the excitation of- and detection of emissions from- fluorophores – fluorescent chemical compounds. With fluorescence microscopy, scientists can observe the location of specific cell types within tissues or molecules within cells.

The difference between the peak of the absorption, or excitation curve and the peak of the emission curve is known as Stoke’s Shift. The greater the distance in this shift, the easier it is to separate the two different wavelengths. Additionally, any overlapping spectrum needs to be removed by the components of the filter cube for reduced background and improved image quality.

As for automatic toys with lasers, Bell says they're just not safe. A curious feline approaching the toy could look right at the light or a sporadic beam could shine into her eye, potentially causing permanent damage.

Exposure of the fluorophore to prolonged excitation will cause it to photobleach, which is a weakening or loss of fluorescence. To reduce photobleaching, you can add an anti-fade mounting medium to the slide and seal the edges with nail polish. The slide should also be kept in the dark when not being imaged.