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Activity 4 Fluorescence Lab Report FINAL

The document discusses fluorescence, a phenomenon where materials emit light of different colors when exposed to UV or blue light. It details a laboratory activity demonstrating fluorescence using highlighters and chlorophyll extracted from spinach leaves, highlighting differences in fluorescence under white and UV light. The document also emphasizes the practical applications of fluorescence in various fields, including security and scientific research.

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8 views6 pages

Activity 4 Fluorescence Lab Report FINAL

The document discusses fluorescence, a phenomenon where materials emit light of different colors when exposed to UV or blue light. It details a laboratory activity demonstrating fluorescence using highlighters and chlorophyll extracted from spinach leaves, highlighting differences in fluorescence under white and UV light. The document also emphasizes the practical applications of fluorescence in various fields, including security and scientific research.

Uploaded by

rdely
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

Names: Bashier Jailani Jr.

, Fevelyn Santillan, Date October 27, 2022


Lovely Joie Fernandez, Regine Mae
Besa, Rey Tristan Dely, Safra Dambong,
Lawrence Faith Coloma, Bench Rawlin
Bautista, Dj Magallanes
Section NO284 Instructo Prof. Rommel Jagus
r

ACTIVITY 4: FLUORESCENCE

Introduction

. Fluorescence is an abrupt emission of electromagnetic radiation because of


excitement happening in a material or substance atom. The absorbed light is different
from the emitted light; it may be weaker because of the energy lost during the process,
and the color may change. However, when the excitement of the molecules is no longer
observed, fluorescence is also switched off. Scientists used fluorescence to detect and
observe molecules that brightly illuminate under UV light (Russell & Endow, 2016).

Fluorescence can be seen in many situations when exposed to ultraviolet (UV) or


blue light. Depending on the circumstances, various organisms or materials absorb UV
or blue light and emit green, yellow, orange, or red light. After the stimulating light is
turned off, the fluorescence is no longer visible. Many plants contain chlorophylls, which
cause them to glow orange or red when exposed to UV or blue light, as well as appear
green in natural sunlight.

Fluorescence is also commonly employed in everyday life for a variety of


purposes, including banknote security to discourage counterfeiting, safety signs and
clothing to boost visibility, and detergents and paper to make them appear whiter. This
laboratory activity demonstrates how fluorescent materials absorb light and releases
light of different color. It also demonstrates that fluorescence lasts only as long as the
fluorescent substance is exposed to the stimulating light.
Results and Discussion

Figure 1. Observing Fluorescence Using Highlighters or Markers

A. fluoresced under the white light. B. fluoresced under UV light.

Figure 2. Observing Fluorescence Produced by Chlorophylls Extracted from Plant


Leaves

A. 3 Medium-Sized Spinach Leaves B. Chlorophyll Under UV Light


To demonstrate fluorescence, the ink from two highlighters was compared, one
of which is fluorescent and the other is not. First, using each of the two markers, a
square is drawn on a transparent plastic sheet and filled in each square with the marker.
The fluorescence was tested by placing a small white light lamp onto the plastic as seen
in figure 1A. There is a slight difference in the color of the two squares where the white
light is focused – the region in the square made by the nonfluorescent marker appears
pale orange, but the region in the square made by the fluorescent marker looks like a
bright shade of orange. To further test the fluorescence of the markers, the transparent
plastic sheet was placed under a UV or black light to view the two squares of orange
ink. There is a noticeable difference between the two squares when viewed under UV
light – the nonfluorescent square appears dark and the fluorescent square fluoresces
light orange (Fig. 1B).
To test the fluorescence of the chlorophyll, the chlorophylls were extracted from a
spinach leaf using a mortar and pestle and about a teaspoon (4-5 ml) of ethanol or
isopropanol (Fig. 2A). The extracted chlorophyll was stored in a test tube with screw cap
then placed in a UV lamp. The chlorophyll fluorescence appears red, as shown in Fig.
2B.

GUIDE QUESTIONS
1. Is there a difference between the squares of yellow made by the two markers
when viewed under UV light?
 Yes
2. If the UV light is turned off, is the green fluorescence still visible?
 When the UV light is turned off, the green fluorescence will not be visible.
3. When exposed to UV light, what color does the plant solution appear?
 Reddish color
4. Why do you think the color is different than when the tube is viewed in room
light?
 It is because light is made up of wavelengths of light, and each
wavelength represents a different color. The color we perceive is caused
by wavelengths being reflected back to our eyes.
5. How is the color of the fluorescence related to that of the UV light that you used
to produce the fluorescence?
 The measured fluorescence colors will vary depending on the substance
and the wavelength of UV light employed.

Understanding and Learning


Upon reading the learning material, there are several lessons and
understandings that have been acquired. It is learned that fluorescence occurs when a
material or substance absorbs light with a given color and then emits another color of
light. Fluorescence is the term used to describe the light that is emitted off of a
substance or material. In other words, fluorescence is the process by which a material
absorbs high-energy light with a short wavelength and releases lower-energy light with
a longer wavelength, usually visible. Fluorescence has a wide range of applications in
modern times. Fluorescence is used by scientists to detect specific chemicals that they
want to investigate further. DNA can be recognized by attaching to substances that are
highly luminous under UV light, for example. Furthermore, fluorescence is widely used
in everyday life for a variety of purposes, including banknote security to prevent
counterfeiting, visibility of safety signs and clothing, and the whiteness of detergents
and paper.

Recommendations/Suggestions/Comments

Mao nalang ni wala. Lihog nalang pud pa check sa spacing if ever.


thankiies
Documentation

References

Colours of light. Science Learning Hub. (n.d.). Retrieved October 27, 2022, from
[Link]
A summary of ultra-violet fluorescent materials relevant to conservation. Australian
Institute for the Conservation of Cultural Material. (2020, August 13). Retrieved
October 27, 2022, from [Link]
fluorescent-materials-relevant-conservation/#:~:text=The%20colours%20of
%20the%20observed,fluoresce%20under%20short%2Dwave%20UV.

Common questions

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Fluorescence in molecular detection relies on principles such as selectivity, sensitivity, and specificity. By using fluorescent tags that bind selectively to target molecules like DNA, researchers can detect these molecules even at low concentrations due to the bright and specific emission under UV light. This high-contrast visibility enhances the ability to study biological processes and biochemical interactions accurately, emphasizing fluorescence's utility in research settings .

Enhancements could include controlled intensity and wavelength of UV light to quantify fluorescence efficiency more accurately. Using spectrophotometers to measure emission spectra would allow precise comparison of fluorescence intensity among samples. Additionally, preparing samples to control concentration and using standardized environments for testing could reduce variability and increase reliability, allowing for broader and more detailed applications such as quantifying pigment concentration in biological studies .

The disappearance of fluorescence when UV light is removed highlights its transient nature, as it ceases immediately when excitation by the external light source stops. This ephemeral characteristic underscores the required continuous energy input for fluorescent emission, as the energy is not stored or prolonged in the fluorescent material, a key principle distinguishing fluorescence from phosphorescence .

Fluorescence demonstrates energy transformation by absorbing light of a higher energy (shorter wavelength) and re-emitting light of a lower energy (longer wavelength). This process occurs because the absorbed energy excites the electrons to a higher energy state, and as they return to their ground state, they release energy in the form of visible light at a longer wavelength. An example from the sources is the chlorophyll which fluoresces red when excited by UV light, indicating this energy transition .

The fluorescent marker emits a bright orange light under UV exposure, whereas the non-fluorescent marker appears dark. This difference reveals that the fluorescent marker's ink contains molecules capable of absorbing UV light and re-emitting it as visible light, while the non-fluorescent ink lacks this property. The chemical structure of fluorescent substances includes conjugated double bonds or specific groups that facilitate this energy transformation, explaining the emission observed .

Fluorescence is used in banknotes to prevent counterfeiting. It involves embedding fluorescent materials that are invisible in normal light but glow under UV light, thereby creating identifiable security features that are difficult for counterfeiters to reproduce. This is because the fluorescent emission is distinct and easily verifiable by banks or other financial institutions .

Fluorescence challenges traditional views by showing that color perception is not solely dependent on ambient light but can be altered by the material properties and absorbed light. It highlights the complexity of light interactions, demonstrating that emitted colors can differ distinctly from absorbed ones, altering perception significantly under different lighting conditions. This introduces a dynamic component to color theory, emphasizing the role of both environmental conditions and molecular properties in perception .

The red fluorescence of chlorophyll under UV light indicates it absorbs the higher energy UV light and re-emits it as lower energy red light. This suggests chlorophyll's specific absorption-emission properties where it preferentially absorbs UV or blue spectrum light and emits at the red end of the visible spectrum, demonstrating its efficiency in energy transfer during photosynthesis .

The choice of fluorescent substances in industrial applications must consider toxicity, environmental impact, cost, stability, and emission properties. Substances that degrade into harmful by-products are less favorable, while those with stable, consistent emission properties under specific lighting conditions are ideal. Additionally, industries prioritize materials that balance cost-effectiveness with functional demands, such as durability and intensity of fluorescence for clear visibility or precise diagnostic applications .

Fluorescence enhances visibility in safety applications by incorporating materials that emit bright light under UV or blue light exposure. In clothing and signs, these materials make objects more visible due to their bright emission, thus increasing awareness and reducing accidents in low-light conditions. This application capitalizes on fluorescence's ability to stand out by emitting vibrant colors against natural backgrounds or in dark settings .

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