Ex vivo confocal imaging using complex micro- scope systems has become a prominent and in- tegral feature of modern-day scientific technol- ogy. Ex vivo conditions enable the process of image acquisition to be optimized and stan- dardized, thus yielding optimal resolution and contrast. The information obtained in this way may be helpful when similar structures have to be identified and interpreted under in vivo con- ditions.
The following examples illustrating blood constituents, pathogenic microorganisms, and ocular tissue structures demonstrate the resolu- tion of the RCM under optimized experimental conditions.
Ex Vivo Applications 4
Fig. 4.1 Ex vivo: Blood, concentrated; rouleaux phe- nomenon
4.1
Blood Components
26 Chapter 4 Ex Vivo Applications
Fig. 4.2 Blood. a–cErythrocytes, different concentrations.dVertical imaging with the Rostock Cornea Module–Heidelberg Retina Angiograph II (TomoCap®)
a b
c d
4.2 Pathogenic Microorganisms 27
Fig. 4.3 Leukocytes: diluted separation
Fig. 4.4 Lymphocytes: diluted separation
a
b
c Fig. 4.5 Acanthamoeba. Ex vivo (a) and in vivo (b) confocal images showing microcysts (cystic stage of life cycle), which are round and up to 10 µm with double-walled structure.cIn vivo confocal image of anterior stroma (depth 93 µm) with several Acan- thamoeba microcysts
4.2
Pathogenic Microorganisms
28 Chapter 4 Ex Vivo Applications
Fig. 4.6 Noncontact confocal microscopy (Nikon
¥50 object lens) of Aspergillus terreus. a Highly reflective conidial heads with conidophores terminat- ing in phialides are visible.bNoncontact confocal
microscopy (Nikon ¥20 object lens) of Aspergillus terreus. The picture shows a cross-section of bran- ched fungal mycelium growing into the agar
a b
Fig. 4.7 Penicillium chrysogenum.a, bWith a non- contact objective system (Nikon ¥50), the culture shows highly reflective brushlike structures at the
level of the agar. These consist of metulae-carrying phialides and conidophores
a b
4.2 Pathogenic Microorganisms 29
Fig. 4.8 Penicillium chrysogenum.aThe picture ob- tained with a noncontact objective system (Nikon
¥20) shows hyaline hyphae growing into the trans-
parent agar.bContact confocal microscopy of highly reflective hyaline hyphae growing into the agar
a b
Fig. 4.9 Reflective opacities in a single stromal plane (depth of focus 180–220 µm) forming branched, needle- like patterns
a b
30 Chapter 4 Ex Vivo Applications
Fig. 4.10 Two-dimensional image and three- dimensional reconstruction of human lens fibers (ex vivo contact mode)
Fig. 4.11 Confocal ex vivo contact microscopy of a rabbit lens.aSurface of the anterior capsular bag (slant image).bLens epithelium on the ante- rior capsular bag with lens fibers.cLens fibers (posterior capsule opacification) after lens refilling with silicon polymer
4.3
Ocular Tissue Structures
a b
c
