Science and educational objects with a world globe floating on a background

Scientists Trot the Globe but Stay Sci-lingual

Academics adapt to new cultures and people, bonding with scientific commonalities.

Image Credit:

©, axllll

When I moved to Germany from India to pursue my masters and graduate studies, the cultural difference was stark. Initially, I was bewildered at cold dinners, succinct chats, and freezing weather, especially coming from a place where warmth is aplenty: in food, weather, and people.

The lab organization differed as well; I almost needed a separate orientation course for the complex reagent labeling system. But just like my other international comrades, I quickly adapted to the local ways, and soon we were pros at lab management and trying glühwein at the Christmas markets. 

My experience is hardly unique. Academia is special; it is perhaps the only profession that normalizes uprooting a life and moving across countries to start afresh every few years. Much like the mutant cells and bacteria we culture in the lab, scientists frequently immerse themselves in new environments, adapt to thrive in them, and emerge stronger from the experiences. 

Travel for academics started as a need when scientific experiments gained popularity. Special equipment and expertise in specific disciplines were rare, so researchers had no choice but to visit research groups across the globe to conduct their experiments. Why is it still a norm today despite the increasing number of world-wide research institutions? Researchers possibly move to work with like-minded experts or to associate with prestigious institutions. But I think that the curiosity of exploring a new place and taking on the associated challenges also contributes to their decisions. 

Since scientists need to always keep open minds and avoid getting stuck in the status quo or traditions, this exercise of acclimatizing to new places, breaking language barriers, and adjusting with diverse people inadvertently broadens their horizons. My hypothesis is that the nomadic life of stereotypically introverted scientists presents a paradoxical situation that pushes them out of their comfort zones in self-experiments that they track throughout their lives. 

How have you navigated moves in your careers?

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Test tubes in laboratory, 96 microwells microplate with ABTS

AI-Powered Automation: Revolutionizing 3D Cell Culture

Researchers streamline cell culture with automated systems, incorporating machine learning to save time and improve reproducibility.

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© adobe

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Scientists create 3D cell culture models such as organoids, spheroids, and organs on a chip for molecular biology research and high throughput drug discovery.1 Unlike costly animal models or 2D cell culture systems that inadequately represent multifaceted tissues, 3D cell culture models are more financially accessible for in-depth biological studies and enable scientists to recapitulate complex physiological functions. However, the culturing process behind models such as organoids is highly involved, often requiring multistep manual methods that are time consuming, laborious, and subject to reproducibility challenges.1

     A 3D rendered image of the Automated Cell Culture System in a laboratory.
The Automated Cell Culture System automates 3D biology, improves workflows, and makes assays more reliable and reproducible.
Molecular Devices

Conventional methods for 3D cell culture rely on human intervention across all steps of growth, including seeding stem cells, collecting aggregates, feeding differentiating cells long term, and imaging and tracking cultures as they grow. Automated cell culture systems that incorporate liquid handling, incubation, monitoring, and imaging optimize these protocols.1

Recent tools that connect automation and artificial intelligence (AI)-mediated feedback systems further optimize this process by modulating culture conditions and screening data with minimal human input.2 For instance, machine-learning algorithms can efficiently monitor and instruct automated technologies for efficient organoid construction, image analysis, and application readouts. AI reduces hands-on time in the laboratory and limits opportunities for human error and variability.1,2

New AI-enabled automation technologies, such as the™ Automated Cell Culture System from Molecular Devices, improve high-throughput cell culture reliability and reproducibility, which is particularly important for 3D culture workflows and drug development.3 This allows researchers to scale up their cell culture models, facilitating faster and more relevant discoveries.

Read more about AI-driven automated cell culture.


  1. Louey A, et al. SLAS Discov. 2021;26(9):1138-47.
  2. Bai L, et al. Bioact Mater. 2024;31:525-48.
  3. Automated Cell Culture System. Molecular Devices. 2023.
A woman is outside with a scent trail behind her that attracts two hungry mosquitoes.

Why Do Mosquitoes Bite Some People More Than Others?

Scientists itch to decipher the cues that make some people mosquito magnets.

Image Credit:

Modified from ©, Rudzhan Nagiev

The incessant buzz of mosquitoes is a sure way to ruin a warm summer evening. Some lucky individuals may escape unscathed, while others become an all-you-can-eat buffet. For those wondering what factors entice hungry female mosquitoes to selectively target someone, scientists have some evidence-driven ideas.

          A picture of a woman with long brown hair wearing a black shirt and brown pants holds a mesh cube with mosquitoes in her hands.
Meg Younger, a neuroscientist at Boston University, studies olfaction in mosquitoes.

A combination of factors, such as carbon dioxide, heat, and skin odor entice mosquitoes. While general cues for warm-blooded animals are carbon dioxide and heat, mosquitoes use odors to distinguish between humans and animals. It turns out that mosquitoes strongly prefer human odors over animal odors.¹

Meg Younger, a neuroscientist at Boston University, noted that carbon dioxide sensitizes mosquitoes to the presence of other odors. “This difference in odor between people accounts for the difference between the attraction of mosquitoes to different people,” she said.

Decoding odor profiles and the corresponding behavioral preferences of mosquitoes is tricky.² “Human odor is composed of hundreds of different volatile chemical molecules that somehow signal the presence of a human,” explained Younger. An emerging theory postulates that the skin microbiome constitutes body odor. “Our skin is covered in bacteria that emit volatiles that vary from person to person.”

While scientists have scratched the surface of quantifying the blend of complex odors that guide mosquitoes, there is good evidence for other factors that entice mosquitoes: being hot and sweaty. Consumption of alcohol, pregnancy, and even malaria infections make a person more likely to be bitten due to increased emitted carbon dioxide and raised body temperatures.³

But don’t worry, mosquito magnets! Scientists hope to develop new repellants to camouflage these tantalizing scents so that pesky mosquitoes can bug off!

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  1. Zhao Z, et al. Nature. 2022;605:706-712.
  2. Giraldo D, et al. Current biology. 2023;33(12): 2367-2382.
  3. Ellwanger JH, et al. Curr Res Parasitol Vector Borne Dis. 2021;1:100058.
An automated pipetting instrument dispenses green liquid into a clear 96-well plate.

Breaking Down Barriers to Single-Cell Resolution

Microfluidic digital dispensing technology can gently isolate viable and healthy cells suitable for a range of downstream applications.

Image Credit:

©, red_moon_rise

Cellular heterogeneity drives many physiological and pathological responses, but conventional analysis methods that sample only in the aggregate can mask signal from rare cell types.1,2 Achieving single-cell resolution has revolutionized scientists’ understanding of cellular behavior, function, and characterization, helping them advance in many fields.2 However, isolating and separating individual cells for downstream single-cell applications can be technically challenging.

     An Uno Single Cell Dispenser instrument, viewed from the side, on a white background.
The Uno Single Cell Dispenser is a benchtop device designed to lower barriers to entry for scientists looking to access single-cell resolution.

Numerous techniques for single-cell separation, isolation, and sorting exist, ranging from manual manipulations to high-throughput instruments capable of processing tens of thousands of cells. In all of these, the primary considerations are yield, quality, purity, throughput, and accessibility.2 Techniques that rely on manual manipulation, such as limiting dilution, do not require sophisticated instruments, but are inefficient and less accurate. Conversely, fluorescence activated cell sorting (FACS) offers high throughputs, but requires complex instrumentation, can be difficult to master, and applies significant mechanical force upon the cells.2

Microfluidics offers a potential third option to this dichotomy. Microfluidic devices separate cells by passing cellular suspensions through microchannels into distinct chambers, thereby providing throughput and accuracy without exerting mechanical stress upon cells.2 While earlier microfluidic devices were complex and highly specialized, newer models are designed with lowering barriers to entry in mind.3 Instruments like the benchtop Uno Single Cell Dispenser™ focus on lower costs, smaller footprints, and greater user-friendliness. None of this comes at the expense of functionality. For example, the Uno can provide 384-well throughput in five minutes with picoliter-level accuracy.Microfluidic digital dispensing technology can gently isolate viable and healthy cells suitable for a range of downstream applications including mass spectrometry-based single-cell analysis, stem cell libraries, 3D cell research, and cell line development.

Read more about microfluidic digital dispensing technology.


  1. Hu P, et al. Front Cell Dev Biol. 2016;4:116
  2. Gross A, et al. Int J Mol Sci. 2015;16(8):16897-919
  3. Sanchez-Avila X, et al. J Am Soc Mass Spectrom. 2023;34(10):2374-80
  4. Uno Single Cell Dispenser™. Tecan Life Sciences. 2023.
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A syringe delivering a fleet of DNA nanorobots to a cancer cell.

Building Biomolecular Machines

William Shih draws inspiration from origami and jigsaw puzzles in his quest to build bigger DNA nanorobots.

Image Credit:

modified from ©, Anastasiia_New, Tilegen, ttsz, Designed by Erin Lemieux

William Shih, a biochemist at Harvard Medical School, explores ways to construct tiny structures out of DNA for biomedical applications. By building on DNA origami, a method for programming DNA to self-assemble into nanoscale shapes, Shih plans to manufacture bigger and more complex molecular robots and to deliver DNA origami cancer vaccines.1  

     Photo of William Shih.
In his lab at Harvard Medical School, William Shih uses structural DNA nanotechnology to design molecular robots for biomedical applications.
Wyss Institute at Harvard University

Why is it challenging to build large structures using traditional DNA origami methods? How did you overcome those challenges?

With DNA origami, we are limited to twice the size of the scaffold strand, which is around 10,000 nucleotides long. It's not feasible to find a scaffold strand long enough to build a larger structure.

We developed an elegant solution called crisscross DNA origami to build the biggest asymmetric structure ever programmed to self assemble—two micrometers by two micrometers—and constructed it from more than 1,000 unique DNA origami components, far surpassing the 64 DNA origami structure that another team built and patterned for the Mona Lisa.2,3 We designed each DNA origami to only interact with specific ports in other DNA origami, akin to a jigsaw puzzle. 

Currently, we can build nanorobots the size of a ribosome, but anything larger, such as a bacterial cell, is difficult. Before crisscross DNA origami, it was hard to imagine how we would do this. Now, we provide a potential route to building larger structures with more sophisticated capabilities.

How do you use DNA origami for therapeutic applications?

Cancer immunotherapy is one of the biggest medical revolutions, but response rates to many life-saving medicines are still low. One reason is that cancer creates an immunosuppressive environment that neutralizes many benefits of these drugs. We developed a DNA origami-based cancer vaccine to help reeducate the immune system to recognize and attack the tumor cells.4 To achieve this, we designed a DNA origami pegboard with tumor antigens on one side and precisely spaced adjuvant oligonucleotides designed to raise the alarm to the immune system attached on the other side. This approach enhanced cytotoxic T cell activation, inhibited tumor growth, and worked synergistically with immunotherapy in a mouse model of melanoma. 

This interview has been edited for length and clarity.


  1. Rothemund PWK. Nature. 2006;440(7082):297-302.
  2. Wintersinger CM, et al. Nat Nanotechnol. 2023;18(3):281-291.
  3. Tikhomirov G, et al. Nature. 2017;552(7683):67-71.
  4. Zeng YC, et al. bioRxiv. 2023.
Image of a floating ghost

Analyzing Phantom Spectra

Katarzyna Tych wants to normalize failure as part of the scientific process.

Image Credit:

©, Paul Campbell

In 2007, I started my graduate studies at the University of Leeds with biophysicist Arwen Pearson and nanoelectronics researcher John Cunningham. I planned to analyze the dynamics of crystallized proteins, which can help us understand how proteins function.

     Kasia Tych wears a grey blazer and smiles at the camera.
Katarzyna (Kasia) Tych earned her PhD at the University of Leeds. She is now a principal investigator at the University of Groningen where she studies macromolecular biophysics.
Blende11, Munich

We wanted to test terahertz time domain spectroscopy, a technique that had never been used to study macromolecular protein crystals, but it had been used to analyze small molecules, for which it produced spectra with sharp, defined peaks. My proteins yielded a lot of sharp peaks, so I thought that things were going well.

But something was wrong. Every time I tried a different sample, the peaks showed up in slightly different positions. We spent more than two years trying to figure out why this kept happening. I finally found a paper with a calculation to determine how much of the spectrum is noise for a given sample thickness and absorption coefficient. It turned out that around 70 percent of our signals—the peaks—were noise; the smooth bits at the beginnings of the curves, which we’d ignored, actually contained the relevant information!

By that time, I was approaching my third year and getting a bit panicky. In my mind, I quit my PhD program maybe 20 or 30 times. I had all kinds of backup plans like opening a cafe with cats and a nice tea selection. But I also had an amazing group of friends; we all vented about our research experiences, so I felt like I was not alone in the suffering.

At the time, it was really difficult. But later I realized how much I had learned from things going wrong. When we embark on research projects, we have to be aware that scientific experiments don’t work a majority of the time. Being aware that failure is normal is incredibly important. We must learn not to say, “I’m a failure because my experiments failed,” but rather, “my experiments failed because that’s just how science works.”

This interview has been edited for length and clarity.

A blue immune cell with a red halo sits in the middle of a yellow spill from a tipping beer mug to the right. Blue bacteria surround the cell.

Alcohol Leaves its Mark on Immune Cells

The immune system's recovery from chronic alcohol use could take longer than some rehabilitation periods due to alcohol's effects on stem cells.

Image Credit:

Modified from ©, Aryo Hadi, anusorn nakdee; designed by ERIN LEMIEUX

Chronic alcohol consumption impairs production of blood and immune cells and dysregulates immune cell functions, but its long term effects remain poorly understood.1-3 Ilhem Messaoudi, an immunologist at the University of Kentucky, and others previously showed that alcohol reduced immune cells’ bacterial killing functions.4,5 In a paper published in Stem Cell Reports, Messaoudi and her team reported that chronic alcohol consumption altered DNA expression in immune stem cells and led to hyperinflammatory monocyte activity.6 These findings suggest that recovering from the effects of alcohol use could take longer than previously anticipated. 

     A schematic showing alcohol’s effects on bone marrow stem cells driving inflammation through changes to DNA accessibility.
Alcohol induces inflammatory changes in immune stem cell progenitors in bone marrow that may last after drinking stops.
Matt Hazard and Thomas Dolan with BioRender

The team first studied a group of rhesus macaques who voluntarily heavily drank alcohol provided to them for one year. After twenty-eight days of abstinence to reflect the shortest period in rehabilitation programs, peripheral blood monocytes from alcohol-drinking animals continued to produce more inflammatory cytokines upon stimulation with bacterial ligands in vitro compared to those from nondrinking animals. 

“That made us think, well, there's got to be a problem with the stem cells,” Messaoudi said. The team isolated bone marrow cells from the animals after one year of heavy drinking and evaluated monocytes and stem cells for gene expression, cytokine production, and epigenetic changes that influence DNA accessibility.

In vitro differentiated progenitor cells from animals that drank heavily skewed toward “neutrophil-like” monocytes with greater inflammatory gene and cytokine expression. These findings matched the chromatin accessibility data, which showed more openness in genes associated with defense and inflammatory responses in bone marrow monocytes and stem cells from alcohol-drinking animals. 

“The complementary techniques they use to analyze the situation were very thorough and very elegant,” said Robert Siggins, a physiologist at Louisiana State University who was not affiliated with the study. 

Messaoudi said that these findings could be important down the road when developing treatments for patients with alcohol use disorders. 


  1. Smith C, et al. Adv Exp Med Biol. 2015;815:349-359
  2. Maraslioglu M, et al. Mediators Inflamm. 2014;2014
  3. Haag F, et al. Eur J Trauma Emerg Surg. 2022;48:2689-2699
  4. Raasch CE, et al. Alcohol: Clin Exp Res. 2010;34(12):2035-2043
  5. Lewis SA, et al. Front Immunol. 2021;12(2021): 724015
  6. Lewis SA, et al. Stem Cell Rep. 2023;18(9):1884-1897
Orange powder in a silver spoon, surrounded by orange pills on a blue background.

Turmeric Tackles Antimicrobial Resistance

An active ingredient in turmeric interacts with light to resensitize pathogens to antibiotics.

Image Credit:

©, ttatty

More than 35,000 people die each year in the US alone from antimicrobial resistant infections.1 With novel treatments in short supply, scientists plan to boost available therapies to target antimicrobial resistance. In a paper published in PNAS, researchers used the power of light to give antibiotics a fighting chance.2

“Everything I do with light is to kill,” said Vanderlei Bagnato, a physicist at Texas A&M University and the University of São Paolo, and coauthor of the paper. 

Light interacts with chemicals called photosensitizers to produce toxic reactive oxygen species.3 Unlike many antibiotics, photosensitizers can sneak past bacterial defenses. Bagnato’s team previously found that light, plus the photosensitizer curcumin, a chemical found in turmeric, potentiated antibiotic efficacy.4   

In their new study, Bagnato and his team tested whether light could resensitize drug-resistant bacteria to lower doses of antibiotics. They found that they needed to expose drug-resistant strains of Staphylococcus aureus to high doses of different antibiotics to curb pathogen growth. However, when Bagnato added the curcumin and shone light on the bacteria, a lower dose sufficed. For most strains, the effects were temporary; the team observed a recurrence in resistance after only a few cycles of bacterial growth. Next, Bagnato wants to dissect the mechanisms underlying the temporary resensitization. 

Tayyaba Hasan, an expert in photomedicine at Harvard Medical School, said that these findings line up with her own findings.3,5 However, she noted that Bagnato and his team only tested gram-positive bacteria. “The real problem is with the gram-negative [bacteria],” said Hasan, since it is harder for antibiotics to breach their outer membrane layers. 

Bagnato emphasized the need for alternative therapies. “Infection does not need one type of weapon; it needs a whole arsenal,” said Bagnato.


  1. CDC. Department of Health and Human Services, CDC;2019
  2. Soares JM, et al. Proc Natl Acad Sci USA. 2023;120(39):e2311667120.
  3. Hamblin MR, Hasan T. Photochem Photobiol Sci. 2004;3(5):436-450.
  4. Soares JM, et al. Sci Rep. 2022;12:21146.
  5. Feng Y, et al. Adv Drug Deliv Rev.  2021;177:113941.
A mouse embryo in which the head, spine, and limb buds are visible.

Illuminating Craniofacial Development

Paul Trainor delves into the genetic and environmental factors that shape the head and face.

Image Credit:

Annita Achilleos and Paul Trainor, Stowers Institute for Medical Research

Craniofacial anomalies are some of the most common birth defects and can severely impact individuals’ lives, potentially compromising the ability to speak, eat, and even breathe. Some of these disorders, known as neurocristopathies, arise from perturbations of the delicate developmental choreography of the neural crest cells. Paul Trainor, a developmental biologist at the Stowers Institute for Medical Research, hopes that determining exactly where, when, and why these cells go awry will inspire new strategies to prevent these disorders.

          Paul Trainor wears a blue shirt and smiles at the camera.
Paul Trainor studies neural crest cells and craniofacial development at the Stowers Institute for Medical Research.
Paul Trainor

What drew you to study neural crest cells?

Neural crest cells are a fascinating population. These cells make most of the bone, cartilage, and connective tissue of the head and face. As they are formed early in embryo development, their proliferation, migration, and differentiation are prone to both genetic and environmental insults. Studying neural crest cells allows me to ask fundamental questions about development, but our research also has potential clinical significance.

How could studying these developmental processes help inform treatments? 

Once we understand the genetic and the molecular processes that cause a specific disorder, we can start to figure out how to correct those processes and prevent the disorder. For example, we know that one disorder, called Treacher Collins syndrome, can be caused by TCOF1 mutations.1 This gene plays a key role in ribosome biogenesis and DNA damage repair. We found that the high levels of oxidative stress naturally experienced by neural crest cells during development can damage DNA; if there are problems with TCOF1, the cell may not be able to repair the damage and will die. In human embryos, we can’t fix the gene, but we might be able to reduce DNA damage. In mice, we showed that antioxidant administration during pregnancy reduced oxidative stress induced DNA damage and increased cell survival, ultimately reducing craniofacial abnormalities.2  

This interview has been edited for length and clarity.


  1. The Treacher Collins Syndrome Collaborative Group. Nat Genet. 1996;12(2):130-136.
  2. Sakai D et al. Nat Commun. 2016;7(1):10328.
Abstract illustration depicting coronavirus research concept.

Curiosity and Compassion Fuel Rare Disease Research

Lauren Drouin shares how personal connections and scientific curiosities drive her work on gene therapy viral vectors. 

Image Credit:

©, DrAfter123

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Lauren Drouin is the director of analytical development and the Genomic Medicine Unit at Alexion AstraZeneca Rare Disease. As a dynamic scientist with unique expertise in current research and industry trends for gene therapies, Drouin is passionate about driving progress within the rare disease field and advancing products from preclinical development into the clinic and beyond.  

In this Science Philosophy in a Flash podcast episode brought to you by Bio-Rad, The Scientist’s Creative Services Team spoke with Drouin to learn more about her interest in adeno-associated virus (AAV) biology, and what motivated her journey from academia to patient-focused analytical development research.

          Headshot of Lauren Drouin.

Lauren Drouin, PhD
Director, Analytical Development
Genomic Medicine
Alexion Pharmaceuticals 
AstraZeneca Rare Disease

Learn more about Lauren Drouin and developing viral vectors.

The image shows different people exercising in a gym.

What Happens to Muscles During Exercise?

Exercise changes our muscles, but its molecular effects depend on the type of exercise. 

Image Credit:

©, vatrushka67

Exercising ranks at the top of many New Year’s resolution lists. While sweating it out, one may wonder what happens inside the body during exercise. According to Keith Baar, a molecular exercise physiologist at the University of California, Davis, the way that people choose to exercise—huffing and puffing on a treadmill or lifting weights—affects how their muscles respond at the molecular level.

          Keith Baar is wearing a patterned shirt. He is standing in front of a tree with his arms crossed.
The molecular exercise physiologist Keith Baar studies how muscles change in response to different types of exercise.
Sasha Dmitriy Bakhter

When people undertake endurance training, such as running, one of the biggest changes in their muscles is an increase in the number of mitochondria, said Baar. “If we go through exercise that uses a lot of energy, our body responds by making more of the machines to make more energy.”   

One key molecule that helps bump up mitochondria numbers is the peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), a protein that binds to transcription factors and regulates the expression of many genes in the skeletal muscles of rodents and humans.1,2 Besides boosting mitochondrial mass,PGC-1α also increases the number of blood vessels in the muscles,4 improving the supply of nutrients and oxygen to keep cells working.

The molecular changes in response to strength exercise, such as lifting weights, are different. “With strength training, we increase the production of the proteins in our muscles by regulating their translation rates,” Baar explained. The mammalian target of rapamycin (mTOR) protein seems to play a central role in muscle growth.5,6 “It regulates anabolic processes, increasing protein synthesis and decreasing protein breakdown,” he said. 

Building both endurance and strength simultaneously may be difficult, though, as the molecular adaptations induced by each type of exercise often counteract one another, Baar explained. “That's why you never see a huge muscular person running a fast marathon.” 

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  1. Baar K, et al. FASEB J. 2002;16(14):1879-1886. 
  2. Mahoney DJ, et al. FASEB J. 2005;19(11):1498-1500.
  3. Wu Z, et al. Cell. 1999;98(1):115-124. 
  4. Chinsomboon J, et al. Proc Natl Acad Sci U S A. 2009;106(50):21401-21406. 
  5. Dreyer HC, et al. J Physiol. 2006;576(Pt 2):613-624. 
  6. Philp A, et al. J Appl Physiol (1985). 2011;110(2):561-568.
A blue immune cell with a red halo sits in the middle of a yellow spill from a tipping beer mug to the right. Blue bacteria surround the cell.

Science Crossword Puzzle

Put on your thinking cap, and take on this fun challenge.

Image Credit:


Click the puzzle for a full-size, interactive version.


1. Variable in a clinical trial, perhaps
4. Output of research 
9. Safety ___ (protective eyewear in the lab)
10. Mature insect stage
11. Exploding star
12. Particle in mass spectrometry
13. Tissue taken in a biopsy, e.g.
15. Period during which the horse genus Harringtonhippus existed 
17. Distress signal at sea
18. Unconfirmed, as a hypothesis
21. Stringed instrument held between the knees
22. ___ tissue (body fat)   
23. Be engulfed by the ocean    
24. Type of population analysis    


1. Breaks down into usable nutrients    
2. Area whose microbiome affects the severity of dandruff    
3. Process in which an embryo leaves the blastula phase    
5. Genus that includes many toxic mushrooms    
6. Fruit of an oak tree    
7. Where Nobel Peace Prizes are awarded    
8. Sucrose, lactose, or maltose    
14. Mixed lettuce salad variety    
16. Like Texas blind salamanders    
17. Neurologist who wrote "Awakenings" and "Seeing Voices"
19. Tidy    
20. Pictures that represent files