How Fluorescence Microscopy Works & How We Save You Time & Money
Despite the diversity in methods, the Excedr lease program can source all instrument types and accommodate any brand preferences your end-user might have. Request an estimate today and see how leasing a TIRF microscope can simplify equipment procurement.
All equipment brands/models are available
The advantages of Excedr’s Fluorescence Microscope Leasing Program:
- Eliminates the upfront cost of purchasing equipment by spreading its cost over time
- Minimizes equipment downtime with included complete repair coverage and preventive maintenance
- Takes advantage of potentially 100% tax deductible* payments, providing you significant cash-savings
- Expedites the administrative work needed for instrument procurement and logistics
- Conserves working capital, enabling you to reinvest in your core business and operations (staffing, inventory, marketing/sales, etc.)
- Accommodates all manufacturer and model preferences
*Please consult your tax advisor to determine the full tax implications of leasing equipment.
Microscopes are found in almost all fields of science, and are used to make small objects more easily visible to the naked eye. Depending on the type, they are used in various areas, from general biology to quantum physics.
Microscopes are organized and grouped based on what method of magnification they use. Optical microscopes, for example, use light and a series of mirrors to produce a magnified image of a sample. The two main categories of optical microscopes are simple and compound.
The difference between simple and compound microscopes is that simple microscopes only use one mirror, while compound microscopes use multiple. The most common optical microscopes use visible light as the light source. However, microscopy utilizes other light spectrums as well.
Fluorescence refers to the emission of light due to the absorption of light. When a material with GFP or other fluorescent dye is exposed to electromagnetic radiation, it absorbs that radiation and re-emits some of it. The time difference, less than a millisecond, between when the organic or inorganic object absorbs the radiation and emits it is short due to the photons’ inherent absorption and emission time.
The object will stop emitting fluorescent light once it stops being hit by radiation. When the materials emit light after the radiation source has gone, it is called phosphorescence. The fluorescent objects’ re-emitted light from the fluorochromes has lower energy and thus a lower wavelength signature, making it exploitable in different analytical techniques.
An example of fluorescence can be seen when one uses a blacklight, and previously unseen colors suddenly are illuminated. Other common examples of autofluorescence include minerals and various bioluminescent animals and plants like algae, fish, and insects.
A fluorescence microscope is a type of light microscope that exploits fluorescence and phosphorescence to identify and observe specific microscopic objects. The area of the material to be observed is dyed with fluorophore tags that will illuminate only the intended parts of the sample.
The specimen is struck with electromagnetic radiation of a specific excitation wavelength that is subsequently absorbed by the fluorophores, which then fluoresce radiation with longer wavelengths than the incident light. This difference in wavelengths can then be used to filter out the light from the light source and only analyze light from the fluorescent sample by using a spectral emission filter.
Detailed imaging of live cells, antibodies, and more can be easily achieved using the fluorescence microscope with proper microscopy techniques. The main components that make up these imaging systems include:
- Light source: Common types are xenon arc lamps and mercury vapor lamps
- Excitation filter: Filters out undesirable specific wavelengths from a light source.
- Dichroic mirror: Filter out specific spectra of light or colors. Also called interference filters or accurate color filters.
- Emission filter: Filters out non-fluorescent light spectra to analyze the desired fluorescence.
Different filter cubes and beam splitters are chosen depending on the specific fluorescent protein used to label the specimen. Single fluorophores are imaged at one time, meaning that a narrow wavelength or one “color” that is emitted from the sample is observed at one time.
High-resolution multi-colored images can be composed by sewing multiple one-color images together. Understanding how to properly perform your fluorescence imaging is critical to fluorescence microscopy since running a sample too many times can cause photobleaching, causing the fluorescent molecules to stop responding.
Fluorophore Imaging Types, Methods, & Cost
The most widely used type of fluorescent microscope is epifluorescence or epi-illumination microscopes.
Most microscopes have light travel from the light source, through the sample, and into the objective lens where the image magnification occurs, deriving the refractive index. This means the light source shines from beneath the sample. In epifluorescence microscopy, the specimen is illuminated from above. This means that the emitted fluorescence and the illuminating light both travel through the same objective lens.
Due to the excitation light traveling through the objective lens to get to the sample and not the other way around, less diffraction occurs overall, meaning less light needs to be filtered out to isolate the fluorescent light.
Total Internal Reflection (TIRF)
A problem that can occur in fluorescence microscopes is that in thicker objects, the light fluorescing from the background matter will drown out the light fluorescing from the object’s surface.
TIRF microscopes are used to focus on just these surface-bound fluorophores’ emitted light. This is done by using evanescent waves for excitation of the fluorophores rather than direct light from a lamp. Evanescent fields or waves are spatially concentrated in the area where they occur.
This means that when these waves occur, they only excite the fluorophores on the sample’s surface. TIRF’s sub-micron surface selectivity makes it the primary method for single-molecule detection due to its specificity.
Super-Resolved Fluorescence Microscopy
For a long time, most of the scientific community accepted the fact that optical microscopy would never be used to observe nano-dimensional objects. The conventional wisdom was that the limitations on the resolution of wavelengths that can be obtained would limit the usefulness of optical microscopy.
The 2014 Nobel Laureates in Chemistry, however, has completely changed this. Eric Betzig, William Moerner, and Stefan Hell were all awarded the Nobel Prize in Chemistry “for the development of super-resolved fluorescence microscopy.” Their discovery of super-resolution microscopy has brought optical microscopy into the nano dimension, making visualization at the 250-nanometer range simple.
Stimulated Emission Depletion Microscopy
In 2000, Stefan Hell developed stimulated emission depletion (STED) microscopy, which utilizes two lasers to achieve nano-level microscopy. One laser is used to fluoresce specific molecules, while the second is used to cancel out undesired wavelengths.
Eric Betzig and William Moerner developed single-molecule microscopy, which allows them to turn molecules’ fluorescent qualities on and off again. Using multiple images with different molecules, either on or off, they can also achieve nano-level microscopy. These are revolutionary techniques that allow scientists to take a better look at the world around us.
Nanoscopy techniques are now being used to look at synaptic behavior between nerve cells in the brain to better understand diseases like Alzheimer’s, Parkinson’s, and Huntington’s disease.
We Offer TIRF Fluorescence Microscope Leases to Fit Every Need
Whatever type of microscope you’re interested in, be it fluorescence, widefield, confocal, electron, or Raman, we’ve got you covered.
If you already have an instrument quote from a manufacturer, you can request a lease estimate below! We’ll start collecting the necessary information to create custom lease terms for you and your lab. If you don’t have a quote yet, get in touch with us above to learn more about what you’ll need to lease.
This off-balance sheet financing structure provides three options at the end of the term. The lessee has the option to return the equipment to the lessor, renew at a discounted rate, or purchase the instrument for the fair market value. Monthly payments are also 100% tax deductible which yields additional monetary savings.
If you recently bought equipment, Excedr can offer you cash for your device and convert your purchase into a long-term rental. This is called a sale leaseback. If you’ve paid for equipment within the last ninety days, we can help you recoup your investment and allow you to make low monthly payments. This also frees up money in your budget rather than tying it down to a fixed asset.
Epifluorescence Microscope Manufacturers & Models on the Market
Light-SHeet: InVi SPIM, QuVi SPIM, MuVi SPIM
BZ-X Series, BZ-X800
FLUOLED Series, FLUOLED TB-SCOPE, FLUOLED Easy, FLUOLED 1CFW, FLUOLED 3CFW, FLUOLED 2CFWMF52,
Epi-fluorescence microscope 158/358
MF43, MZX11, MZX81, MF53, MF31, MF52-N, MF31-UV, MF31-RB
EVOS Series, EVOS M5000, EVOS M7000, EVOS XL Core, EVOS FL, EVOS FL Auto, EVOS FL Auto 2, EVOS FLoid, Cellnsight CX7, Cellnsight CX5
Axio Zoom.V16, Axio Scan.Z1, Celldiscoverer 7, Lightsheet Z.1, Apotome.2
MVX10, IXplore Pro, IXplore Standard, BX63, BX53
M205 FCA, M205 FA, M165 FC, MZ10 F, TCS SP8 STED
Lionheart LX, Cytation 1
iScope, Oxion, Oxion Inverso, bScope, B+ series, DZ series, Delphi-X Observer
LW Scientific:>br/> Innovation Lumin, i4 Lumin
View Solutions Inc.