Application of confocal laser scanning microscopy to the study of amber bioinclusions

Abstract

Confocal laser scanning microscopy is an essential analytical tool in biological, biomedical, and material sciences, integrating microscope manufacturing technology, optical-electronic technology, and computer technology. In the last decade, confocal laser scanning microscopy has been successfully applied to the study of amber bioinclusions. Enhanced signal to noise ratios, resolution power, capability of optical sectioning, three-dimensional reconstruction, and better performance when imaging thicker samples provide a great deal of valuable and detailed morphological information about amber fossils. We briefly discuss the practical applications of CLSM in amber studies and compare it with other imaging methods commonly used in the field, including bright-field microscopy, wide-field fluorescence microscopy, and micro-computed tomography. A general procedure for imaging amber inclusions with CLSM is provided, with a focus on pretreatments and image processing.

New species of Omma Newman from mid-Cretaceous Burmese amber (Coleoptera, Archostemata, Ommatidae)

“…1B). As such, we think O. liassicum cannot be confidently differentiated from Beutelius (and even some more distantly related genera, e.g., Bukhkalius Kirejtshuk & Jarzembowski;Li et al 2021). Kirejtshuk (2020) created the genus Cionocups to accommodate a new species from Burmese amber, Cionocups manukyani.…”

“…Widefield fluorescence images were mainly captured with a Zeiss Axio Imager 2 light microscope combined with a fluorescence imaging system. Confocal images were obtained with a Zeiss LSM710 confocal laser scanning microscope, using the 488 nm Argon laser excitation line (Fu et al 2021) Etymology. The specific name is from the Latin 'fortis', meaning strong, referring to the robust appearance of the species.…”

A New Species of Vetubrachypsectra from Mid-Cretaceous Amber of Northern Myanmar (Coleoptera: Brachypsectridae)

“…Confocal images were obtained with a Zeiss LSM710 confocal laser scanning microscope, using the 488 nm (Argon; for V. huchengi sp. nov.) or 561 nm (DPSS 561-10; for V. burmitica) laser excitation lines [20]. Fluorescence images could illustrate the structures better than brightfield ones, as the fluorescence emitted around the border between the inclusion and amber matrix clearly shows the boundary of structures.…”

On the Nosodendridae from mid-Cretaceous amber of northern Myanmar (Coleoptera: Nosodendroidea)

“…Photographs under incident light were mainly taken with a Zeiss Discovery V20 stereo microscope. Confocal images were obtained with a Zeiss LSM710 confocal laser scanning microscope, using the 488 nm Argon laser excitation line (Fu et al 2021). Images under incident light and widefield fluorescence were stacked in Helicon Focus 7.0.2 or Zerene Stacker 1.04.…”

“…16C). Since the strong laser could penetrate the somewhat opaque amber matrix better, and out-of-focus light is blocked, the confocal microscope could image this material much better than widefield ones (Fu et al 2021). The medial antennomeres of B. pengweii appears to be relative elongate (Fig.…”