Torsdagskvällsreflektioner + 1 artikel

Tjenixen alla PHI,are. Förmodligen tycker ni som jag att aktiehandeln och kursen är miserabel.
Mig veterligen finns det nada fundamentamässigt som föranlett denna utveckling.
Förmodligen är det (förhoppningsvis) de sista skvättarna ur garanternas innehav som kör ner aktien.
Som ex idag, någon som använder sig av Penser (EPB) avyttrade 4000 aktier till nästan vilket pris som helst. Men men, snart borde dessa garanter vara ute ur aktien så vi får mer normal handel baserad på Bolagets utveckling och förknippade händelser.
Under tiden fortsätter bloggen sitt grävarbete oförtröttligt (nåja).

Här nedan kommer PHI-relaterad info.
Bloggen skrev i augusti om en forskningsrapport som nyligen hade släppts. Inlägget.
Innehållet var som vanligt intressant och visade på HoloMonitorn´s användbarhet i forskning även utanför de traditionella som ex cancerforskning.
Jo, forskarna bakom rapporten har nu sammanställt text som berättar mer i detalj hur och med vilka tekniker de använde för att nå sina resultat och fått en artikel utgiven i branschtidningen Micron.
Volume 113, October 2018, Pages 1-9

Micron

The International Research and Review Journal for Microscopy

Micron is an interdisciplinary forum for all work that involves new applications of microscopy or where advanced microscopy plays a central role. The journal will publish on the design, methods, application, practice or theory of microscopy and microanalysis, including reports on optical, electron-beam, X-ray microtomography, and scanning-probe systems. It also aims at the regular publication of review papers, short communications, as well as thematic issues on contemporary developments in microscopy and microanalysis. The journal embraces original research in which microscopy has contributed significantly to knowledge in biology, life science, nanoscience and nanotechnology, materials science and engineering.

I Oktoberutgåvan av tidskriften finns nu mer info om DHM tekniken och fylligare info m exempelbilder från PHI`s HoloMonitor.Jag saxar valda delar ur artikeln:

- The recently developed digital holographic microscopy (DHM) has enabled the quantitative evaluation of cell size and morphology without labeling or destruction.

- Therefore, DHM was validated as a promising device for testing platelet function given that it allows for the quantitative evaluation of activation-dependent morphological changes in platelets. DHM technology will be applicable to the quality assurance of platelet concentrates, as well as diagnosis and drug discovery related to platelet functions.

- Recent developments and subsequent advancements in digital holographic microscopy (DHM) have enabled the continuous or intermittent observation of morphological changes in the same cells with regard to cell height, volume, surface roughness, surface irregularity, and many other indexes without destruction, fixation, or labeling . In general, morphological changes in cells in vitro are microscopically examined using selected views. Consequently, given the limited number of cells subjected to qualitative evaluation, data obtained therefrom may not necessarily represent the overall trend in cell populations. DHM, however, allows the scanning of more than 1000 cells within minutes, as well as the quantitative evaluation of their morphology.
Therefore, it has the advantage of being able to detect small but statistically significant differences among similar cell populations.

- Compared with aggregometry or flow cytometry (FCM), which have typically been used for evaluating platelet activation, we demonstrated the major advantages of DHM in the simultaneous quantitative analysis of adherent single and aggregated platelets without staining or labeling

Data acquisition and DHM image analysis

Fixed platelets on plastic culture wares were directly subjected to imaging without labeling or staining using a digital holographic microscope (HoloMonitor M4; Phase Holographic Imaging AB, Lund, Sweden). As essentially described in a previous study regarding human periosteal cells and mesenchymal stem cells (Kawase et al., 2016), images were captured and analyzed using a specifically designed software (HoloStudio M4; Phase Holographic Imaging AB). In each sample, more than five snapshots of randomly selected views were taken to obtain a total of at least 1500 platelets. All images were binarized, and platelets were segmented under the same conditions (adaptive mean = 128; object size = 8) using the manual mode. Acquired data (N = 1800) were then plotted in histograms using the aforementioned software. Alternatively, quantitative data were exported to Excel (Microsoft, Redmond, WA, USA) for further statistical processing as described in the following section.

Given that the aforementioned software adopts “1.38” as the default setting for the cell refractive index as a default value by the manufacturer (Phase Holographic Imaging AB) (Kawase et al., 2016) based on previously published data (Beuthan et al., 1996; Rappaz et al., 2008; Yu et al., 2009; Persson et al., 2010; Svet, 2013), this value was utilized in the determination of optical thickness and platelet volume. The refractive index of the surrounding medium had been fixed at 1.34 (default) throughout the experiment (Kawase et al., 2016).

- The same samples were then observed by DHM. DHM hologram images for time-course changes in platelet appearance are shown in Fig. 5. These images were further converted to 3D reconstruction images (Fig. 6). As shown in a 2D hologram image (Fig. 5), white small round particles grew in size (i.e., area), whereas those particles decreased, with time of treatment. On the other hand, 3D reconstruction imaging (Fig. 6) has an advantage for the comparison of height (i.e., thickness). The number of tall platelet aggregates apparently increased with time of treatment.

Fig. 5. DHM hologram images for time-course changes in platelet appearance. As described in the legend of Fig. 2, washed platelets were prepared and treated without (A) or with 0.1% CaCl₂ for (B) 5 min, (C) 10 min, or (D) 20 min. After treatments, platelets were fixed and subjected to DHM examination. The magnification for all images remained constant (Bar = 100 μm). Similar data were obtained from three additional experiments using different donor samples.

Fig. 6. DHM 3D reconstruction images for time-course changes in platelet appearance. As described in the legend of Fig. 2, washed platelets were prepared and treated without (A) or with 0.1% CaCl₂ for (B) 5 min, (C) 10 min, or (D) 20 min. After treatments, platelets were fixed and subjected to DHM examination. The magnification for all images remained constant (Bar = 100 μm). Similar data were obtained from three additional experiments using different donor samples.

The present study tested the feasibility of DHM examination in the quantitative evaluation of morphological changes in activated platelets. Accordingly, DHM is able to examine both living and fixed cells without further labeling or staining. Although fixed cells are suitable for comparing multiple groups at once, the advantage of using living cells is that DHM may be employed to continuously monitor changes in platelet morphology. Considering that the present study needed to simultaneously compare multiple platelet populations obtained from the same blood sample, we opted to use fixed platelets rather than living ones.

During DHM examination, we demonstrated that platelets activated using CaCl₂ shifted from the lower left to the upper right region. Moreover, FCM analysis demonstrated that CaCl₂ upregulated CD62P expression in single platelets, while SEM examination demonstrated that single platelets aggregated in response to CaCl₂ stimulation. Although other platelet function tests had not been performed to further confirm our observations, the amount of data obtained herein is sufficient to suggest that the apparent region shift in the two-dimensional DHM data plots mainly represents the aggregation of activated platelets.

In conclusion, we herein attempted to validate the applicability of DHM in the quantitative evaluation of activated platelets. Data obtained from our feasibility tests implied that DHM could be a promising method for quantitatively examining morphological changes in platelets in vitro.

Regarding clinical relevance, we indicate that this technology will be further applicable during laboratory testing. To date, we have vigorously investigated the quality and potency of platelet concentrates in regenerative dentistry and reached a conclusion that a quality check of individual PRP preparation is indispensable for predictable PRP therapy (Kawase and Okuda, 2018). At present, the methodology regarding a quick and sensitive quality check of platelets is a major matter remaining to be developed. We believe that DHM will be a promising device for this purpose. Furthermore, probably more importantly and influentially in medical fields, DHM will be applied in the diagnosis of platelet disorders and drug discovery for controlling platelet functions as a high throughput system.

Min kommentar

Läser man igenom hela artikeln får man känslan av att de i minsta detalj skärskådat PHI`s teknik (DHM) och letat svagheter o annat som kan vara till teknikens nackdel.

Men och just ett stort MEN, de kommer istället till slutsatsen : 

"Data obtained from our feasibility tests implied that DHM could be a promising method for quantitatively examining morphological changes in platelets in vitro.

Regarding clinical relevance, we indicate that this technology will be further applicable during laboratory testing.

We believe that DHM will be a promising device for this purpose. Furthermore, probably more importantly and influentially in medical fields, DHM will be applied in the diagnosis of platelet disorders and drug discovery for controlling platelet functions as a high throughput system."

Om nu någon aktieägare (baserat på aktiens nuvarande handel) tvivlar på teknikens förträfflighet och kommande kommersiella genombrott, 11 japanska forskare vibbar om dess raka motsats.

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