Det handlar om studier kring ett nytt ämne att använda inom fluorescerande mikroskopi användbart exempelvis vid diagnos och terapi för cancerpatienter.
Synthesis of a dihalogenated pyridinyl silicon rhodamine for mitochondrial imaging by a halogen dance rearrangement
Received 31 May 2019, Accepted 05 Sep 2019, Published 01 Oct 2019
Abstract
Results: A dihalogenated fluorinatable pyridinyl rhodamine could be successfully synthesized with the high yield of 85% by application of a halogen dance (HD) rearrangement. The near-infrared dye shows a quantum yield of 0.34, comparable to other organelle targeting SiR derivatives and absorbs at 665 nm (εmax = 34 000 M−1cm−1) and emits at 681 nm (τ = 1.9 ns). Using colocalization experiments with MitoTracker® Green FM, we could prove the intrinsic targeting ability to mitochondria in two human cell lines (Pearson coefficient >0.8). The dye is suitable for live cell STED nanoscopy imaging and shows a nontoxic profile which makes it an appropriate candidate for medical imaging.
Conclusions: We present a biocompatible, nontoxic, small molecule near-infrared dye with the option of subsequent radiolabelling and excellent optical properties for medical and bioimaging. As a compound with intrinsic mitochondria targeting ability, the radiolabelled analogue can be applied in multimodal (PET/OI) imaging of mitochondria for diagnostic and therapeutic use in, e.g., cancer patients.
Värt att notera är att i denna studie har man använt sig av STED (STimulated Emission Depletion) mikroskopteknik för att få sanslöst högupplösta bilder. Som ex denna:
Varför är det intressant och vad har det för koppling till HoloMonitor frågar sig den skeptiske.
Jo,av 2 anledningar.
1. STED-mikroskopets uppfinnare Stefan W. Hell fick som ni kanske minns Nobelpriset för detta 2014.
Två år senare köper Stefan Hells forskningsavdelning en HoloMonitor.
Lab of Nobel Price winner purchases HoloMonitor
The Department of Optical Nanoscopy (Prof. Stefan W. Hell) at the German Cancer Research Center (DKFZ) in Heidelberg recently ordered a motorized HoloMonitor M4. Professor Hell was awarded the Nobel Prize in Chemistry in 2014 for his pioneering work in the field of super-resolution fluorescence microscopy. The customer list price of the ordered unit is 350 000 SEK.
“It is a great honor for us to have the research team
of a Nobel Prize winning scientist added to our growing list of
customers, selecting our HoloMonitor microscope for their research
needs”, said CEO Peter Egelberg.
2. Den nu aktuella studien är utförd av Stefan Hells forskningsavdelning, DKFZ, och man har "trumvirvel" använt sig av HoloMonitor för att få fram sina resultat.
Urklipp från studien. (fetningar i texten är bloggens)
Although for PET examinations only nano- or picomolar amounts of the
radiopharmaceutical compound are needed, medical applications of
fluorescence dyes (e.g., fluorescence-guided interventions) require
larger amounts of material. Therefore, cytotoxicity testing is necessary
for our proposed bimodal imaging agent 16 and its precursor 15. For toxicity assessment, the frequency and duration of cell division with and without incubation with dye 15 was analyzed via time-lapse holographic imaging (Figure 3).
U2OS cells were incubated with 1 µM dye 15
in medium for 1 h and, after washing with dye free medium, continually
imaged over a period of 14.5 h using a holographic incubator microscope.
The analysis of the data revealed that the frequency and duration of
cell division of the cells incubated with dye 15 show no difference to the untreated control (frequency of cell division with SiR dye 15: 0.30 ± 0.05 divisions per cell, without dye: 0.32 ± 0.06 divisions per cell, for division duration see Figure S7, Supporting Information File 1).
These results are supported by cell count and confluency analysis (Figure S9, Supporting Information File 1).
In summary, we conclude that dye 15
does not show any significant cytotoxicity in this human cell line.
Comparable experiments with HeLa cells strengthen these results (data
not shown).
Figure 3:
Exemplary holographic image sequence of two cell divisions of U2OS cells treated with 1 µM of dye 15. Dividing cells round up and can be distinguished from non-dividing cells by height. After incubation with 1 µM dye 15 for 1 h, the cells were washed and then holographically imaged using a HoloMonitor®
M4 time-lapse cytometer. Cell proliferation was followed for 14.5 h (30
min between images) and corresponding time-lapse movies are available
in Supporting Information Files 2–5, scale bar 50 µm.
Och ur den kompletterande filen
6. Assessment of cytotoxicity
Potential cytotoxicity of SiR dye 15 was assessed via analyzing duration and frequency of cell division using holographic time-lapse imaging with a HoloMonitor®M4 cytometer.
One day prior to the experiment, U2OS (human bone osteosarcoma epithelial) cells were seeded in a 24-well cell culture plate.The medium of one half of the wells was replaced with medium containing 1μM SiR dye 15 and the other half with fresh medium without dye. Incubation was done for 1h at 37°C and 5% CO2.
All wells were washed three times with medium before holographic time-lapse imaging was started using PHI HoloLidsTM to ensure optimal image quality.
Images were recorded for 14.5h with one image of each well every 30min.
Data analysis was done with the HstudioM4 Software including cell segmentation, tracking of dividing cells (Figure S7), cell counting and confluency measurement (Figure S8).
De forskningsrapporter som nu kommer in där HoloMonitor använts bygger på tidigare kedja av studier där man inte haft tillgång till detta excellenta instrument.")
Här får vi ett konkret bevis för hur användbar HoloMonitor är, i detta fall som kontrollinstrument till det superavancerade STED-instrumentet. Wiki kan berätta mer om STED.
Min kommentar
Att Nobelpristagaren Stefan W. Hells forskningsavdelning kommit fram till de resultat denna studie visar, att man lyckats ta fram ett helt nytt ämne till fromma för cancerdrabbade, och det tillsammans med PHI`s instrument HoloMonitor är en stor framgång för Bolaget.
Detta tippar bloggen kommer användas i Bolagets marknadsföring.
Ska vi gissa att denna studie får branschens bjässar att kika närmare på vad detta är för instrument och PHI som dess ägare är?
Ja,förutom de 2 jänkare som Bolaget redan förhandlar med. Dessa 2 friare får nog sockra sina förslag efter denna studie. VD sitter i en jäkligt bra sits. Måtte han ha is i magen och alla aktieägare tålamod.
Mvh the(euforiska)99
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