How Light Sheet Fluorescence Microscopy Works & How We Save You Time & Money

Excedr’s lease program can source all instrument types and accommodate any brand preferences your end-user might have despite the diversity in types. Request a light sheet fluorescence microscope lease estimate today and see how leasing your next light sheet microscope can simplify equipment procurement and budget management.

All equipment brands/models are available

The Advantages of Excedr’s Light Sheet Fluorescence Microscope (LSFM) 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.

a microscope, an ocular lens, and microscopic image next to each other

Light sheet microscopy is essential when imaging large, sensitive, living specimens that are easily damaged by light exposure.

As a fluorescence imaging technique, it utilizes a planar sheet of light in order to illuminate the live sample and excite fluorophores within a specific focal volume. The excitation light can either be created using a cylindrical lens or virtually generated using a scanning beam or dithering beam array.

The latter technique is normally referred to as digitally scanned laser light sheet microscopy (DSLM), while the former is called selective plane illumination microscopy, or SPIM.

Detection occurs along an axis perpendicular to that of illumination so that only the section being observed is lit, rather than the entire specimen or any out-of-focus features. This planar illumination technique produces optical sectioning, or clear images of multiple focal planes within the sample.

Furthermore, using the perpendicular axis reduces the amount of photo-damage, photo-bleaching, and phototoxicity to the specimen so that imaging can occur long-term.

It makes light sheet microscopy more ideal compared to processes like wide-field fluorescence microscopy that can damage your sample more rapidly and reduce long-term viability.

Optical sectioning offers a wide field of view, increased contrast, faster image acquisition speeds, and high axial resolution and spatial resolution. These capabilities support a variety of applications, including the real-time, three-dimensional imaging monitoring of neural responses in live organisms, and enable the 3D reconstruction of a 2D image by computationally combining the image data from a stack of images.

Light sheet microscopy’s principle benefits apply across multiple fields, including drug discovery, neurobiology, developmental biology, embryology, plant biology, and more.

Being able to 3D image everything from large, live organisms, tissue explants, to 3D cell cultures and single cells is crucial in many different areas of research.

This is especially true in developmental biology, where many model organisms are used to understand human development, such as fruit fly embryos (Drosophila melanogaster), an excellent model system for understanding the basic biology underlying our own embryonic development.

Another example of these organisms is the zebrafish, which is primarily used to study vascular development and disease.

Light sheet imaging has been pivotal in 3D examination, avoiding disruption or damage of the organism’s development cycle.

Methods, Configurations, & Light Sheet Microscope Price

It is first important to mention that there is no perfectly planar light sheet. The sheet is only approximated and given over a range of values using different methods such as static planar sheets or scanned beams that approximate a light sheet over time. Some examples of these methods include:

Planar

This optical imaging technique is relatively older, and uses a singular cylindrical lens to project the light sheet into the sample, allowing illumination to be expanded across a single axis. This is also known as selective plane illumination microscopy or SPIM.

Compared to a traditional confocal microscope, acquisition rates using this method are much faster by several orders of magnitude.

Scanned

Also called a digital sheet, this principle involves scanning a light sheet in lateral directions and then stitching the images together. This creates a larger composite 3D image, and is beneficial for observing moving samples.

The downside of this method is that it scans in both the axial and lateral directions, meaning longer acquisition times and more exposure to light.

Multiphoton

Both of the aforementioned techniques can be used in addition to multiphoton excitation. Two photon light sheet microscopy uses a rare phenomenon where two photons are absorbed in a single quantum event. This happens at longer wavelengths that are optimal for imaging deep tissues in high resolution.

This type of excitation coupled with Bessel beam/airy beam light sheets helps reduce unfavorable aspects of those illumination patterns, including out-of-focus excitation/illumination.

Bessel Beam/Airy Beam

This technique was first developed by the lab of Alexander Rohrbach. Bessel beams are used to create digitally scanned light sheets and are formed by shaping the illumination light with an axicon, a specialized type of lens which has a conical surface.

A Bessel beam is a non-diffracting wave that does not “spread out” as light usually does. If a Bessel beam is obstructed, it can “self-heal” and reform further down the axis. These aspects combine to enable enhanced imaging at greater depths within biological specimens. Airy beams function in the same vein, as they are also both non-propagating and “self-healing.”

Lattice

This type of light sheet fluorescence microscopy is used to accurately observe live cells for longer periods of time without affecting or damaging their behavior. It allows for high speed acquisition and subcellular resolution through the use of a structured light pattern rather than a thicker, non-uniform sheet of light.

It uses the same fundamentals as a Bessel beam microscope, but aims to reduce illumination out of the depth of field of the specimen by creating a lattice of Bessel beams.

Image Processing

LSFM has become the technique of choice to image dynamic processes Because of reduced light dosage and, hence, lower phototoxicity and photobleaching.

To properly analyze complex dynamics within these datasets, image quality has to be improved, either during acquisition or through post-processing, to remove artifacts coming from stripe patterns or to increase signal contrast and penetration depth.

Stripe Pattern Removal

Because LSFM datasets have excellent axial resolution, dynamic range, and low signal to noise, there don’t generally require any preprocessing routines. However, there are limits to image quality.

One of them includes stripe patterns that exist due to the absorption of light on the surface of the sample. The stripes create shadows that impede the use of segmentation tools used to track various cell characteristics.

There are different approaches to handle stripe pattern removal. One examples include post-acquisition processing tools, such as variational stationary noise removal or multiview fusion, which allow the correction of artifacts up to some extent. These methods do require powerful workstations and long processing times, however.

Another example is a double-sided illumination system, which allows researchers to pivot the light sheet illumination angle and create longer integration times at the camera to obtain an average image, remove stripes from shadows during acquisition, and reduce image processing time.

Multiview Reconstruction & Deconvolution

Sample rotation is one of the main characteristics of light sheet fluorescence microscopy techniques.

It allows researchers to observe large specimens with high resolution from different view angles, avoiding harmful effects that sample absorption and scattering introduce. Using sample rotation, biological features that would otherwise be hidden are observed.

Furthermore, sample rotation is exceptionally helpful on single-sided illumination setups, where a face of the sample is poorly illuminated.

Light Sheet Fluorescence Microscope Leases to Fit Every Need

an ocular lens and eyepiece tube with a microscopic image displayed

Need to acquire a LSFM for your living specimen and cell imaging needs? The upfront costs of a light sheet fluorescence microscope may be higher than you are able or willing to pay. If that’s the case, you’re probably looking for an alternative to purchasing the microscope in order to create more flexibility within your budget.

Whatever your budgetary goals, when you lease a light sheet fluorescence microscope with Excedr, you can deploy the capital you do save on upfront costs to other areas of your business, such as hiring, buying consumables and reagents, or expanding your lab space.

Because our approval process is up to 12 weeks faster than traditional financing methods, you can acquire the equipment much quicker, accelerating your R&D or drug development and hitting milestones sooner.

If you have a quote from a manufacturer, we can put together a lease estimate for you. Or, if you’d just like to learn more about our leasing program, get in touch with us today.

Furthermore, we offer confocal microscopy leasing options in case you’re considering light sheet microscopy alternatives for imaging biological samples. If you’re unsure of whether a light sheet microscope or confocal microscope will suit your needs best, check out our confocal microscopy page.

Operating 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.

Sale-Leaseback

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.

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