Laser Scanning Multiphoton Microscopes
It is fully script-capable and connects via an application pro- gramming interface (API) and a dynamic link library (DLL) to external programs or programming languages like Matlab or Python. It also handles all interfaces to control scanning and a variety of image analysis platforms, including ImageJ. Multi- photon scanning is implemented via the widely known and very powerful MBF Vidrio ScanImage soſtware.
Integration with Original Equipment Manufacturer (OEM) Systems Te vertical integration capability of the MPX-1040 micro-
scope frame allows integration into OEM applications where standard confocal scanning systems have reached their limit in terms of penetration depth and optical sectioning perfor- mance. Compact, highly integrated, and industry-qualified packaging enables the world’s first multiphoton scan engine to be integrated as an OEM component in advanced high-content screening or molecular cell biology applications.
Multimodal Microscopy in Medical Research and Nose-to-Brain (N2B) Drug Delivery Microscopy techniques are crucial in
the field of medicine, especially for diag- nosis of diseased tissue. Currently, the workflow for diagnosis from a biopsy or resection is tissue fixation followed by par- affin sectioning and histological staining, for example, hematoxylin and eosin (H&E) staining. Tis is time-consuming, requires experienced technicians for the staining, and includes toxic staining reagents, which are huge drawbacks in the standard pro- tocols. Te use of label-free biomarkers in pathology would provide faster and easier diagnosis. Figure 5 shows unlabeled FFPE sections of a melanoma (A) and a lung tumor (B) where autofluorescence in the widefield and multiphoton mode, as well as SHG signal originating from the collagen, shows clear differences in the appearance of the cancerous compared to normal tissue. Te delivery of drugs to the central
nervous system (CNS) through the intra- nasal pathway is a very important topic in pharmaceutical research, especially for the growing number of patients suffering from neurological disorders. It is critical to ensure the treatment compound, for exam- ple, antibodies, proteins, and derivatives, can selectively and accurately reach its pharmacological target, and safely achieve the desired therapeutic effect. Te so-called “nose-to-brain” (N2B Patch Project: www.
n2b-patch.eu) approach provides a robust and promising route for drug transporta- tion while bypassing the restrictive blood brain barrier (BBB) [9]. Tis is a big advan- tage when compared with commonly used methods,
for example, intrathecal 2022 May •
www.microscopy-today.com
or intracerebroventricular methods. Tis project successfully achieved the specific targeting of the olfactory, or the respiratory, region, by using a catheter-based refined technique [9]. Figure 6A shows a widefield-fluorescence WSI of a sagit-
tal cryosection of a mouse head. Te WSI image and regions of interest depicted by Figures 6B–6F were acquired with the MPX- 1040, upright configuration, with fully automated slide load- ing (up to 200 slides at a time). Te sagittal view of the murine head gives an overview of key regions of interest in the olfac- tory region, that is, respiratory mucosa, olfactory mucosa with adjacent olfactory bulb, anterior olfactory nucleus, and parts of the brain. Te slides were scanned using an xyz-stage and an in- house-developed soſtware routine. Macro-imaging (Figure 6A) was achieved using a 4× lens (Torlabs, Super Apochromatic objective, Air, NA 0.2) and three excitation channels (λex/em): blue (395/432 nm), green (475/515 nm) and red (637/681 nm). Higher-magnification images (Figures 6B–6F) were collected using a 16× objective (Nikon LWD Plan Fluorite, water, NA 0.8) for microimaging for more detailed analysis.
Figure 5: Label-free widefield (A) and multiphoton (B) WSI of human skin melanoma (A) and human lung cancer (B) FFPE sections. A) Autofluorescence of human melanoma collected by widefield fluorescence (λex/ λem 390/432 nm; magenta), 475/515 nm (green) and 555/595 nm (orange). Sample kindly provided by Dr. Branislav Zagrapan, LKFH Feldkirch - AT. B) Autofluorescence of human lung tumor collected by multiphoton imaging at 1040 nm excitation SHG (magenta), 542 nm (green), 595 nm (yellow). Sample kindly provided by Dr. Lucas Onder, KSSG
St.Gallen - CH. Scale bars = 1 mm.
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