Rhodamine Synthesis Essay

This lab could have contained errors. The errors could have happened when performing the lab. Some of the possible errors in this lab are:

Abstract: The purpose of this experiment is to use the apparatus shown in figure 7-1 of experiment 6 found in the lab manual, to synthesize [1,3,5-C6H3(CH3)3]Mo(CO)3.(2016 Oleg) With this, characterization of the molecule can be accomplished using the infrared spectrum and NMR spectrum of the synthesized compound. It was found in the IR spectra of the product, that suitable stretches were found associated with the C-O bonds of mestylene and molybdenum. With One strong spectra was found at 1855.3973 cm-1 , one medium strength spectra was found at 1942.6002 cm-1 , and one weak spectra of C-O stretch was found at 1298.8638cm-1 . The 1H NMR spectrum of the product showed peaks at δ 2.28 and 5.25, while the H NMR spectrum of mesitylene gave peaks at δ

[2] McCray, J. A., & Trentham, D. R. (1989). Properties and uses of photoreactive caged compounds. Annual review of biophysics and biophysical chemistry,18(1), 239-270.

The goal of this experiment is to prepare a photosensitive solution and explore its properties. While analyzing the solution, one will learn how to successfully handle these sensitive chemicals and then establish its properties via spectrophotometry.

The aim of this investigation was to determine the effect of ethanol on the membrane permeability using Beta vulgaris. Beta vulgaris contains red pigments called betalain sequestered in vacuoles. The cell membrane is generally impermeable to betalain as this pigment is relatively large and cannot pass through the membrane by diffusion. (123HelpMe.com, 2015) However, by increasing the permeability of the cell membrane, betalain can leach out of the cell and colour the liquid red. The colour intensity of the solution due to leakage of betalain is proportional to the membrane permeability. To quantify the colour intensity, the light absorbance of the solutions containing a Beta vulgaris cube were measured by a spectrophotometer. These measurements were used to analyse the membrane permeability. (Flinders University, 2015)

Evers, D. J., B. Hendricks, G. Lucassen, and T. Ruers. "Optical Spectroscopy: Current Advances and Future Applications in Cancer Diagnostics and Therapy." NCBI. National Center for Biotechnology Information, 8 Mar. 2012. Web. 13 Sept. 2015.

The concentrations of L-arginine and L-asparagine were measured in the range 0.110 mM. Data was collected from the fluorescence intensity of the Arg-and Asn-sensing membranes at the two emission wavelengths (λem = 565 and 625 nm) at an excitation wavelength of 460 nm (λex = 460 nm). The fluorescence spectra for detecting L-Arg and L-Asn were measured using a multifunctional fluorescence microtiter plate reader (Saphire2, Tecan GmbH, Wien, Austria).

This complete process is visualized under a microscope by feeding the Tetrahymena cells different concentrations of India Ink and then observing vacuole formation. The purpose of this experiment was to test if the concentration of India ink affects the rate of vacuole formation. To discover this, we tested 3 Tetrahymen mixed with a 1%, 2%, and 3% and counted how many vacuoles were absorbed. If the India Ink concentration is changed, then there will be a change in the formation of

Fluorescent properties are used to study protein and small molecule interactions. Fluorescent spectrophotometry determined the excitation wavelength of eosin isothiocyanate to be 525 nm and its emission wavelength was 545 nm. Glycogen phosphorylase was similarly studied. The excitation wavelength was 330 nm and the emission wavelength was 360 nm. The emission wavelength could be indicative of the presence of the fluorophore, tryptophan. The potential interaction between bovine serum albumin (BSA) and 1-anilino-8-naphthalene sulfonic acid (ANS) was studied through fluorescence and fluorescence resonance energy transfer (FRET). BSA had an emission wavelength of 358 nm which could also be indicative of a tryptophan. When BSA and ANS were mixed together, the emission wavelength was longer suggesting that the molecules interact with each other and follow FRET.

Rhodamine B appears green in powder form but when added to water turns a vivid fluorescent pink.

In this experiment, the use of a glass cuvette was used in order to avoid any chemical or environmental interference from the surroundings. Furthermore such glass cuvettes can support large wavelengths during transmission of light beyond 320 nm since this is the excitation wavelength region. Riboflavin is excited at around 370 nm, therefore the wavelength range must be large in order to detect a proper response. Plastic cuvettes are however inadequate as the range of wavelengths supported for transmission of light is very limited (5, Upstone). The geometry of the instrument of the spectrophotofluorometer has a specific angle and position for the excitation source and the detector. The excitation polychromator emits ultra-violet radiation and light to the interested sample and the shape of the polychromator resembles a triangular prism in which the monochromator has been angled at a 90∘ position. The transducer would detect the irradiation excitation beam and transfers it through the emission polychromator. The emitted beam would then travel to the detector to allow for the response signal to appear for the sample on the computer system (416, Skoog). The position of the cuvette is essential as it enables for the consistent readings of the response signals, as any shift in the cuvette position can create variation in results due to the position of lighting and detection of molecular samples hitting at different angles. The process of fluorescence is quite sensitive than absorption due to having lower detection limits because of the fact that it can detect lone molecules in excited states that are irradiated by UV-light unless they come into contact with other molecules within

First; a brief introduction about the confocal microscopy, critical aspects of confocal Microscopy, and the basic parts of the confocal microscope. It will follow by more information about fluorescence microscopy which is used for study fixed and living cells because of its versatility, specificity, and high sensitivity.

Direct optical imaging methods offer several benefits, namely a swift acquisition time, relatively low market price and high spatial resolution. The cost and image resolution will go up if microscopic imaging techniques are to be used. However, there are some disadvantageous using this technique. For example, with increasing imaging resolution, the field of view will decrease. One solution is to scan the contamination plume, but at the price of reduced temporal resolution. In general, the resolution of optical imaging is restricted by the light intensity, magnification and the pixel density of the camera. Moreover, image acquisition time is affected by the light intensity and the amount of light penetrating through the boundaries of the sample. Often the rate of exposure needs to be lowered in samples with low photon density.

However, under cellular imaging conditions, Spinach and Broccoli show greatly reduced fluorescence compared with nominal brightness. It has been known that the fluorescence decrease of Spinach−DFHBI is not due to irreversible photobleaching like GFP. It is caused by reversible conversion to a non- fluorescent state and the signal can be recovered

For cellular biology analyses, I have two years of training in confocal microscopy to perform complex experiments including: Fluorescence Resonance Energy Transfer, Fluorescence Recovery After Photobleaching, real time live cell imaging. These provided the means to analyze both the spatial and temporal properties of interacting proteins using microscopy. Cell