MORONE DIEGO (IT)
| US20140121493A1 | 2014-05-01 | |||
| US20200104998A1 | 2020-04-02 |
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| CLAIMS 1. A rigid skinfold chamber element provided with at least two ((1) and (2)) and optionally at least a third (3) fixation points, an optical window (4) surrounded by a surface from which at least 4 arms (5) ,(6), (7) and (8) protrude whereby two substantially identical elements can be held together by suitable fixation means at the fixation points in order for the skinfold chamber to delimit an area of skin which can be examined through the optical window. 2. The element of claims 1, wherein the fixations systems (1) and (2) are located on the short side of the rectangle circumscribing such element. 3. The element of claim 2, wherein the fixation systems (1) and (2) are located on two arms (5) and (6) of the element that protrude in opposite directions. 4. The element of claim 3, wherein the third fixation element (3) is located on the long side of the rectangle circumscribing such element. |
This invention pertains to a rigid skinfold chamber which features allow its use without surgical intervention. The skinfold chamber of the invention provides adequate exposure and stabilisation of the skin area to be studied, and is particularly useful for the study of tissue damage and repair, tumor growth and angiogenesis.
BACKGROUND
Transparent chambers for the study microcirculation date as back as 1930s, when they were used mainly for the investigation of microcirculation in rodents. In 1993 Lehr et al. published a seminal paper in which dorsal skin-fold chamber was applied to nude mice. Thereafter, different skin- fold chambers have been employed for the investigation of tumor angiogenesis, cell migration and metastasis, thrombosis, volume therapy and biocompatibility of engineered tissues. These studies all rely upon the same skin-fold chamber design as Lehr et al., (for example those described in Alieva et al., 2014 or Alexander et al., 2008) which require the surgical exposure of a dorsal skin flap and the suture of a glass-protected chamber for the visualization of the subcutaneous and partially of the dermis layers of the skin. This surgical intervention implies additional animal stress and suffering, limits the possibility to perform subsequent acquisitions over time, and most importantly alters any measurement associated with the cellular metabolic changes of the skin area being investigated. There is thus a need to develop skinfold chambers which do not require surgical implantation of the same.
BRIEF DESCRIPTION OF THE FIGURES
Fig. 1. Skinfold chamber of the invention
Fig. 2. Example of a skinfold chamber according to the invention
Fig. 3. Example of use of a skinfold chamber according to the invention
DETAILED DESCRIPTION OF THE INVENTION.
We have designed a skinfold chamber which removes the need for surgical implantation of the device, provides adequate exposure and stabilisation of the skin area to be studied, and is particularly useful for the study of tissue damage and repair, tumor growth and angiogenesis.
With reference to Fig. 1, this inventions provides a rigid skinfold chamber element provided with at least two ((1) and (2)) and optionally at least a third (3) fixation points, an optical window (4) surrounded by a surface from which at least 4 arms (5), (6), (7) and (8) protrude whereby two substantially identical elements can be held together by suitable fixation means at the fixation points in order for the skinfold chamber to delimit an area of skin which can be examined through the optical window.
In one embodiment, the fixations systems (1) and (2) are located on the short side of the rectangle circumscribing such element.
In another embodiment, the fixation systems (1) and (2) are located on two arms (5) and (6) of the element that protrude in opposite directions.
In a more specific embodiment, the third fixation element (3) is located on the long side of the rectangle circumscribing such element.
The invention is now described by reference to the figures and examples below.
Figure 1: Skinfold chamber of the invention. A skinfold chamber of the invention highlighting the following features: fixation points (1), (2) and (3), optical chamber (4), protruding arms (5), (6), (7) and (8)
Figure 2. Example of a skinfold chamber according to the invention , and which constitutes a specific embodiment of the invention. (A) the chamber shown from the exterior (left) and interior (right) part, with screws and bolts for mounting. The chamber has an imaging window of 10 mm. To further preserve the sample, a coverslip of size 11 mm can be mounted and fixed with an O-ring of same size. (B) schematics of the chamber with dimensions. (C) mounted chamber.
In this example, the skinfold chamber is made in brass, which does not make it suitable for use in Magnetic Resonance Imaging (MRI). The skilled artisan will be able to choose , from the plethora of rigid materials available to him, other suitable rigid materials such as a non-metallic polymeric rigid material in order to obtain a skinfold chamber according to the invention that is compatible with the instrumentation used to observe the skin area of interest.
Figure 3. Example of use of a skinfold chamber according to the invention. (A) chamber is mounted on the mouse by pinching the back skin with forceps and enclosing the wound inside the chamber window. (B) side view of the mounted chamber. ( C) intravital stage consists of a Plexiglas base, a thin heating pad, paper and a XYZ micrometric manipulator. (D) L-shaped stainless-steel bracket is mounted on the XYZ manipulator. Skinfold chamber can be fixed on the bracket with plasticine or an additional screw. (E) fixation of the skinfold chamber on the intravital stage. (F) side view of the intravital stage on the microscope table. (G) detail of the mounted imaging window. (H) corresponding diagram with two possible imaging grids embracing wound bed and perilesional skin. (I) skinfold chamber can be unmounted after imaging without damaging the skin.
REFERENCES
Alexander, S., Koehl, G.E., Hirschberg, M., Geissler, E.K. & Friedl, P. Dynamic imaging of cancer growth and invasion: a modified skin-fold chamber model. Histochem Cell Biol 130, 1147-1154 (2008).
Alieva, M., Ritsma, L., Giedt, R.J., Weissleder, R. & van Rheenen, J. Imaging windows for long-term intravital imaging: General overview and technical insights. Intravital 3, e29917 (2014).
Lehr et al, Am J Pathol, Oct; 143(4): 1055-62. (1993).